Collagenase inhibitor

ABSTRACT

Compounds having inhibitory activity against inhibitory activity against type IV collagenase and are useful as angiogenesis, cancer infiltration or cancer metastasis inhibitors. The compounds have the formula: ##STR1## in which R 1  represents a group of formula: --OR 3  (wherein R 3  represents a hydrogen atom or an alkyl group), 
     --NR 4  R 5  (wherein R 4  and R 5  each represents a hydrogen atom, an alkyl or alkoxy group), 
     --NHCH(R 6  COR 7  (wherein R 6  represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, and R 7  represents an alkyl group), 
     --NHCH(R 6 )COOR 8  (wherein R 8  represents an alkyl group) or 
     --NHCH(R 6 )CONR 9  R 10  (wherein R 9  and R 10  each represents a hydrogen atom or an alkyl group, or NR 9  R 10  together represent a heterocyclic ring group); and 
     R 2  represents a hydrogen atom, an alkyl or aralkyl group.

This application is a 371 of PCT/JP92/01420 filed Nov. 2, 1992.

TECHNICAL FIELD

The present invention relates to novel piperazic acid derivatives having excellent collagenase inhibiting activity.

BACKGROUND ART

Collagenase degrades collagen, which is one of the main components of the connective tissues. Of the collagenases, type IV collagenases degrade type IV collagen, which is a main component of the basement membranes. It is known that the activity of type IV collagenases is raised at the time of angiogenesis accompanied by cancer growth and at the time of cancer invasion and metastasis, and that these enzymes play an important role in the degradation of the basement membranes [William G. Stetler-Stevenson; Cancer and Metastasis Reviews, Vol. 9, 289-303 (1990)]. Accordingly, a collagenase inhibitor may be useful for the prevention and treatment of these diseases.

Hitherto, there have been reports concerning low molecular weight substances showing collagenase inhibiting effects, including peptide compounds containing a mercapto group [Robert D. Gray, Hossain H. Saneii and Arno F. Spatola: Biochemical and Biophysical Research Communications, Vol. 101, No. 4, 1251-1258 (1981); Charles F. Vencill, David Rasnick, Katherine V. Crumley, Norikazu Nishino and James C. Powers: Biochemistry, Vol. 24, 3149-3157 (1985)]; peptide compounds containing a carboxyl group [Jean-Marie Delaisse, Yves Eeckhout, Christopher Sear, Alan Galloway, Keith McCullagh and Gilbert Vaes: Biochemical and Biophysical Research Communications, Vol. 133, 483-490 (1985)]; benzyloxycarbonyl propyl-leucyl-glycylhydroxamic acid [William M. Moore and Curtis A. Spilburg: Biochemical and Biophysical Research Communications, Vol. 136, 390-395 (1986)]; and hydroxylamine derivatives (Japanese Patent Kokai Application Hei No. 1-160997). Furthermore, SC 44463 has been reported [Reuven Reich, Erik W. Thompson, Yukihide Iwamoto, George R. Martin, James R. Deason, George C. Fuller and Ruth Miskin: Cancer Research, Vol. 48, 3307-3312 (1988)] as an inhibitor relatively specific to type IV collagenases. SC 44463 has been confirmed to exhibit an inhibitory activity on cancer metastasis in an animal experiment. The above compounds, however, are synthetic ones and are not yet put to practical use.

On the other hand, the tissue inhibitor of metalloproteinase (TIMP) and related substances are known as collagenase inhibitors of the protein type. It becomes possible to prepare TIMP on a large scale by recombinant DNA technology, but it has not yet been put to practical use [A. J. P. Docherty, A. Lyons, B. J. Smith, E. M. Wright, P. E. Stephens, T. J. R. Harris, G. Murghy and J. J. Reynolds: Sequence of human tissue inhibitor of metalloproteinases and its identity to erythroid-potentiating activity; Nature, Vol. 318, 66-69 (1985)].

As a compound having a hydroxylamino-substituted 2-pentylsuccinic acid structure, actinonin has been isolated from the culture filtrate of actinomycetes. This compound has been reported to inhibit aminopeptidase M at low concentrations [H. Umezawa, T. Aoyagi, T. Tanaka, T. Suda, A. Okuyama, H. Naganawa, M. Hamada and T. Takeuchi: J. Antibiotics, Vol. 38, 1629-1630 (1985)], however, it has not been investigated whether or not it inhibits type IV collagenases.

Furthermore, a natural product having the following chemical structure has been isolated from a strain which belongs to streptomyces and is known to have antibacterial and collagenase inhibiting activities (Japanese Patent Kokai Application Hei 3-157372). ##STR2##

In addition, in Japanese Patent Kokai Application Hei 3-53891, the formula ##STR3## of the compound is described, and the compound is said to have antibacterial activity, however, there is no disclosure of its inhibitory activity on type IV collagenases.

DISCLOSURE OF INVENTION

The present inventors have eagerly studied the synthesis of derivatives having better inhibitory activity against type IV collagenases and the pharmacological activities of these derivatives. The study resulted in the finding that new matlystatin derivatives have excellent inhibitory activity against type IV collagenases and that these derivatives can be useful inhibitors of angiogenesis, inhibitors of cancer invasion and inhibitors of cancer metastasis; and they completed the present invention.

CONSTITUTION OF THE INVENTION

The compounds of the present invention have the general formula: ##STR4##

In the above formula, R¹ represents a group of formula: --OR³ (wherein R³ represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms), --NR⁴ R⁵ (wherein R⁴ and R⁵ are the same or different and each represents a hydrogen atom, an alkyl group containing 1 to 4 carbon atoms or an alkoxy group containing 1 to 4 carbon atoms), --NHCH(R⁶)COR⁷ (wherein R⁶ represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms and R⁷ represents an alkyl group containing 1 to 4 carbon atoms), --NHCH(R⁶)COOR⁸ (wherein R⁶ is as defined above and R⁸ represents an alkyl group containing 1 to 4 carbon atoms) or --NHCH(R⁶)CONR⁹ R¹⁰ (wherein R⁶ is as defined above, and R⁹ and R¹⁰ are the same or different and each represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms, or NR⁹ R¹⁰ together represent a heterocyclic ring group); and R² represents a hydrogen atom, an alkyl group containing 3 to 16 carbon atoms or an aralkyl group comprising an optionally substituted phenyl group and an alkyl group containing 1 to 4 carbon atoms.

However, those compounds in which R¹ represents a group of formula: --NHCH(R^(6a)) COR^(7a) (wherein the combination of R^(6a) and R^(7a) signifies the isobutyl and methyl groups or the ethyl and sec-butyl groups) and R² represents a pentyl group are excluded.

In the said formula (1), the term "alkyl" used in connection with an alkyl group containing 1 to 4 carbon atoms represented by R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹ and R¹⁰ falling under the definition of R¹ an alkyl moiety of an alkoxy group containing 1 to 4 carbon atoms represented by R⁴ and R⁵ and an alkyl moiety containing 1 to 4 carbon atoms of an aralkyl group falling under the definition of R² signifies a straight or branched chain alkyl group containing 1 to 4 carbon atoms, such as a methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl group; preferably, in the case of R⁶ falling under the definition of R¹, a methyl isopropyl isobutyl or sec-butyl group, and, in the case of the other R's, an alkyl group containing 1 or 2 carbon atoms.

In the said formula (1), where R² represents an aralkyl group, examples of the substituents on the phenyl moiety include an alkyl group such as a methyl or ethyl group, an alkoxy group such as a methoxy or ethoxy group, and a halogen atom such as chlorine or bromine.

In the said formula (1), where R² represents an aralkyl group, preferred examples of the complete aralkyl group include an unsubstituted benzyl or phenethyl group.

In the said formula (1), where R¹ represents a heterocyclic ring group represented by NR⁹ R¹⁰ together, examples of such heterocyclic ring groups include nitrogen-containing heterocyclic groups such as pyrrolidin-1-yl, imidazolin-1-yl, pyrazolidin-1-yl, pyrazolin-1-yl, piperidino, piperazin-1-yl and morpholino groups preferably pyrrolidin-1-yl, piperidino and morpholino groups.

In the said formula. (1), where R² represents an alkyl group containing 3 to 16 carbon atoms, it signifies a straight or branched chain alkyl group such as an n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, n-heptyl, 1-methylhexyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 1-propylbutyl, 4,4-dimethylpentyl, n-octyl, 1-methylheptyl, 2-methylheptyl, 3-methylheptyl, 4-methylheptyl, 5-methylheptyl, 6-methylheptyl, 1-propylpentyl, 2-ethylhexyl, 5,5-dimethylhexyl, n-nonyl, 3-methyloctyl, 4-methyloctyl, 5-methyloctyl, 6-methyloctyl, 1-propylhexyl, 2-ethylheptyl, 6,6-dimethylheptyl, n-decyl, 1-methylnonyl, 3-methylnonyl, 8-methylnonyl, 3-ethyloctyl, 3,7-dimethyloctyl, 7,7-dimethyloctyl, undecyl, 4,8-dimethylnonyl, dodecyl, tridecyl, tetradecyl, pentadecyl, 3,7,11-trimethyldodecyl, hexadecyl, 4,8,12-trimethyltridecyl, 1-methylpentadecyl, 14-methylpentadecyl or 13,13-dimethyltetradecyl group; preferably an alkyl group containing 4 to 12 carbon atoms; and more preferably an alkyl group containing 6 to 10 carbon atoms.

Preferred compounds are those in which:

2)

R¹ represents a group of formula:

--OR³ (wherein R³ represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms);

--NR⁴ R⁵ (wherein R⁴ and R⁵ are the same or different and each represents a hydrogen atom, an alkyl group containing 1 to 4 carbon atoms or an alkoxy group containing 1 to 4 carbon atoms);

--NHCH(R⁶)COR⁷ (wherein R⁶ represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms, and R⁷ represents an alkyl group containing 1 to 4 carbon atoms); or

--NHCH(R⁶)CONR⁹ R¹⁰ (wherein R⁶ is as defined above, and R⁹ and R¹⁰ are the same or different and each represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms, or NR⁹ R¹⁰ together represents a heterocyclic ring group); and

R² represents a hydrogen atom, an alkyl group containing 3 to 16 carbon atoms or an aralkyl group comprising an optionally substituted phenyl group and an alkyl group Containing 1 to 4 carbon atoms;

3)

R¹ represents a group of formula:

--OR³ (wherein R³ represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms);

--NR⁴ R⁵ (wherein R⁴ and R⁵ are the same or different and each represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms); or

--NHCH(R⁶)COR⁷ (wherein R⁶ represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms, and R⁷ represents an alkyl group containing 1 to 4 carbon atoms); and

R² represents a hydrogen atom, an alkyl group containing 3 to 16 carbon atoms or an aralkyl group comprising an optionally substituted phenyl group and an alkyl group containing 1 to 4 carbon atoms;

4)

R¹ represents a group of formula: --NR⁴ R⁵ (wherein R⁴ and R⁵ are the same or different and each represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms); and

R² represents an alkyl group containing 3 to 16 carbon atoms or an aralkyl group comprising an optionally substituted phenyl group and an alkyl group containing 1 or 2 carbon atoms; and

5)

R¹ represents a group of formula: --NR⁴ R⁵ (wherein R⁴ and R⁵ are the same or different and each represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms); and

R² represents an alkyl group containing 6 to 10 carbon atoms.

The compounds of formula (1) in accordance with the invention can exist in various stereoisomeric forms due to the presence of asymmetric carbon atoms having the R- and/or S-configuration. Although all of the isomers are represented herein by a single formula, the present invention covers not only mixtures of the isomers but also each of the individual isomers.

Examples of the compounds of the invention are listed in Table 1. Such examples are not to be construed as being limitative of the invention.

In Table 1 the abbreviations used have the following significance.

Me: methyl group;

Et: ethyl group;

Pr: propyl group;

iPr: isopropyl group;

sBu: sec-butyl group;

iBu: isobutyl group;

Pen: pentyl group;

Hex: hexyl group;

Hep: heptyl group;

Oct: octyl group;

Dec: decyl group;

Ph: phenyl group;

PhE: phenethyl group;

Am: amyl group;

Pyrd: pyrrolidin-1-yl group;

Imid: imidazolidin-1-yl group;

Pyzr: pyrazolidin-1-yl group;

Pyz: pyrazolin-1-yl group;

Pipe: piperidino group;

Pipr: piperazin-1-yl group; and

Mor: morpholino group.

                  TABLE 1                                                          ______________________________________                                          ##STR5##                                                                                  Compound                                                           Compound No.  R.sup.1            R.sup.2                                       ______________________________________                                          1            OMe                Pen                                            2            OMe                iBu                                            3            OMe                Oct                                            4            OMe                H                                              5            OtBu               Pen                                            6            OtBu               iBu                                            7            OtBu               Oct                                            8            OtBu               Hex                                            9            OtBu               Hep                                           10            OtBu               Dec                                           11            OtBu               PhE                                           12            OtBu               H                                             13            HNMe               Pen                                           14            HNMe               iBu                                           15            HNMe               Oct                                           16            HNMe               Hex                                           17            HNMe               Hep                                           18            HNMe               Dec                                           19            HNMe               PhE                                           20            HNMe               Et                                            21            HNMe               Me                                            22            HNMe               Pr                                            23            HNMe               Bu                                            24            N(Me).sub.2        Pen                                           25            N(Me).sub.2        iBu                                           26            N(Me).sub.2        Oct                                           27            N(Me).sub.2        Hex                                           28            N(Me).sub.2        Hep                                           29            N(Me).sub.2        Dec                                           30            N(Me).sub.2        PhE                                           31            N(Me).sub.2        Et                                            32            N(Me).sub.2        H                                             33            N(Me)OMe           Pen                                           34            N(Me)OMe           iBu                                           35            N(Me)OMe           Oct                                           36            N(Me)OMe           Et                                            37            N(Me)OMe           H                                             38            NHOMe              Pen                                           39            NHOMe              iBu                                           40            NHOMe              Oct                                           41            NHOMe              Et                                            42            NHOMe              H                                             43            NHPh               Pen                                           44            NHPh               iBu                                           45            NHPh               Oct                                           46            NHPh               Et                                            47            NHPh               H                                             48            NHCH.sub.2 Ph      Pen                                           49            NHCH.sub.2 Ph      iBu                                           50            NHiBu              Pen                                           51            NHiBu              iBu                                           52            NHiAm              Pen                                           53            NHiAm              iBu                                           54            HNCH(iPr)COEt      Pen                                           55            HNCH(iPr)COEt      iBu                                           56            HNCH(iPr)COEt      Oct                                           57            HNCH(iPr)COEt      Et                                            58            HNCH(iPr)COEt      H                                             60            HNCH(iBu)COEt      iBu                                           61            HNCH(iBu)COEt      Oct                                           62            HNCH(iBu)COEt      Et                                            63            HNCH(iBu)COEt      H                                             64            HNCH(sBu)COEt      iBu                                           65            HNCH(sBu)COEt      Oct                                           66            HNCH(sBu)COEt      Et                                            67            HNCH(sBu)COEt      H                                             68            HNCH(iPr)COOMe     Pen                                           69            HNCH(iPr)COOMe     iBu                                           70            HNCH(iPr)COOMe     Oct                                           71            HNCH(iPr)COOMe     Et                                            72            HNCH(iPr)COOMe     H                                             73            HNCH(iPr)CONHMe    Pen                                           74            HNCH(iPr)CONHMe    iBu                                           75            HNCH(iPr)CONHMe    Oct                                           76            HNCH(iPr)CONHMe    Et                                            77            HNCH(iPr)CONHMe    H                                             78            HNCH(iPr)COOtBu    Pen                                           79            HNCH(iPr)COOtBu    iBu                                           80            HNCH(iPr)COOtBu    Oct                                           81            HNCH(iPr)COOtBu    H                                             82            HNCH(iPr)CON(Me)OMe                                                                               Pen                                           83            HNCH(iPr)CON(Me)OMe                                                                               iBu                                           84            HNCH(iPr)CON(Me)OMe                                                                               Oct                                           85            HNCH(iPr)CON(Me)OMe                                                                               H                                             86            HNCH.sub.2 COOMe   Pen                                           87            HNCH.sub.2 COOMe   iBu                                           88            HNCH.sub.2 COOMe   Oct                                           89            HNCH.sub.2 CONHMe  Pen                                           90            HNCH.sub.2 CONHMe  iBu                                           91            HNCH.sub.2 CONHMe  Oct                                           92            HNCH.sub.2 CON(Me).sub.2                                                                          Pen                                           93            HNCH.sub.2 CON(Me).sub.2                                                                          iBu                                           94            HNCH.sub.2 CON(Me).sub.2                                                                          Oct                                           95            HNCH(Me)COOMe      Pen                                           96            HNCH(Me)COOMe      iBu                                           97            HNCH(Me)COOMe      Oct                                           98            HNCH(Me)CONHMe     Pen                                           99            HNCH(Me)CONHMe     iBu                                           100           HNCH(Me)CONHMe     Oct                                           101           N(Me)CH.sub.2 COOMe                                                                               Pen                                           102           N(Me)CH.sub.2 COOMe                                                                               iBu                                           103           N(Me)CH.sub.2 COOMe                                                                               Oct                                           104           N(Me)CH.sub.2 CONHMe                                                                              Pen                                           105           N(Me)CH.sub.2 CONHMe                                                                              iBu                                           106           N(Me)CH.sub.2 CONHMe                                                                              Oct                                           107           HNCH(Ph)COOMe      Pen                                           108           HNCH(Ph)COOMe      iBu                                           109           HNCH(Ph)COOMe      Oct                                           110           HNCH(Ph)CONHMe     Pen                                           111           HNCH(Ph)CONHMe     iBu                                           112           HNCH(Ph)CONHMe     Oct                                           113           HNCH(CH.sub.2 Ph)COOMe                                                                            Pen                                           114           HNCH(CH.sub.2 Ph)COOMe                                                                            iBu                                           115           HNCH(CH.sub.2 Ph)COOMe                                                                            Oct                                           116           HNCH(CH.sub.2 Ph)CONHMe                                                                           Pen                                           117           HNCH(CH.sub.2 Ph)CONHMe                                                                           iBu                                           118           HNCH(CH.sub.2 Ph)CONHMe                                                                           Oct                                           119           HNCH(iPr)CONH.sub.2                                                                               Pen                                           120           HNCH(iPr)CONH.sub.2                                                                               iBu                                           121           HNCH(iPr)CONH.sub.2                                                                               Oct                                           122           HN-c-Hx            Pen                                           123           HN-c-Hx            Oct                                           124           HN-c-Hx            iBu                                           125           HNCH(CH.sub.2 OMe)COOMe                                                                           Pen                                           126           HNCH(CH.sub.2 OMe)COOMe                                                                           iBu                                           127           HNCH(CH.sub.2 OMe)COOMe                                                                           Oct                                           128           HNCH.sub.2 CONH.sub.2                                                                             Pen                                           129           HNCH.sub.2 CONH.sub.2                                                                             iBu                                           130           HNCH.sub.2 CONH.sub.2                                                                             Oct                                           131           HNMe               H                                             132           HNMe               H                                             133           N(Me)OMe           H                                             134           NHOMe              H                                             135           NHPh               H                                             136           Pyrd               Pen                                           137           Pyrd               iBu                                           138           Pyrd               Oct                                           139           Pyrd               Hex                                           140           Pyrd               Hep                                           141           Pyrd               Dec                                           142           Imid               Pen                                           143           Imid               iBu                                           144           Imid               Oct                                           145           Imid               Hex                                           146           Imid               Hep                                           147           Imid               Dec                                           148           Pyzr               Pen                                           149           Pyzr               iBu                                           150           Pyzr               Oct                                           151           Pyzr               Hex                                           152           Pyzr               Hep                                           153           Pyzr               Dec                                           154           Pyz                Pen                                           155           Pyz                iBu                                           156           Pyz                Oct                                           157           Pyz                Hex                                           158           Pyz                Hep                                           159           Pyz                Dec                                           160           Pipe               Pen                                           161           Pipe               iBu                                           162           Pipe               Oct                                           163           Pipe               Hex                                           164           Pipe               Hep                                           165           Pipe               Dec                                           166           Pipr               Pen                                           167           Pipr               iBu                                           168           Pipr               Oct                                           169           Pipr               Hex                                           170           Pipr               Hep                                           171           Pipr               Dec                                           172           Mor                Pen                                           173           Mor                iBu                                           174           Mor                Oct                                           175           Mor                Hex                                           176           Mor                Hep                                           177           Mor                Dec                                           ______________________________________                                    

Of the compounds illustrated in the above table, preferred compounds are Compounds No. 1, 2, 3, 4, 5, 6, 7, 13, 14, 15, 16, 17, 18, 19, 23, 24, 25, 26, 27, 28, 29, 30, 33, 34, 35, 54, 55, 56, 60, 61, 62, 64, 65, 66, 67, 68, 69, 73, 74, 82, 83, 86, 87, 88, 89, 90, 91 95, 101, 104 and 107. The more preferred compounds are Compounds No. 1, 2, 3, 5, 6, 13, 14, 15, 16, 17, 18, 19, 24, 26, 27, 28, 29, 30, 54, 55, 56, 60 and 64. The most preferred compounds are:

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N-methylamide,

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid N-methylamide,

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid N-methylamide,

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N-methylamide,

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid N-methylamide,

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid N-methylamide,

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N,N-dimethylamide,

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid N,N-dimethylamide,

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid N,N-dimethylamide,

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N,N-dimethylamide,

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid N,N-dimethylamide, and

N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid N,N-dimethylamide.

The compounds of the invention can be prepared by the procedure summarized in the following scheme.

That is, the compounds of formula (1) of the invention can be prepared by 1 reacting a compound (a), which is a key intermediate, with a compound (b) in the presence of a condensing agent to prepare a compound (c); 2 removal of a protecting group, B⁴, of the compound (c), to prepare a compound (d); 3 reacting a compound (d) with an alcohol or amine, R¹ H, followed by removal of the protecting group, B⁴, after which the product is reacted with a hydroxylamine, B⁶ ONH₂, to prepare a compound (e); and 4 removal of protecting groups, A and B⁶. ##STR6##

In the formula of the above reaction scheme, R¹ and R² are as defined above and the significance of A, B¹, B⁴ and B⁶ will be explained later. Furthermore, methods for the preparation of the compounds of the invention will be explained in detail. ##STR7##

In the formulae of the above reaction scheme, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹ and R¹⁰ are as defined above; A represents an amino-protecting group (preferably a benzyloxycarbonyl group); B¹ represents a carboxyl-protecting group (preferably a tert-butyl or benzyl group); B² represents a carboxyl-protecting group (preferably a methyl or ethyl group); B³ represents a tri-substituted silyl group (preferably a trimethylsilyl group); B⁴ represents a carboxyl-protecting group (preferably a benzyl, tert-butyl, trichloroethyl or tribromoethyl group); B⁵ represents an amino-protecting group (preferably a benzyloxycarbonyl, tert-butoxycarbonyl, allyloxycarbonyl or trichloroethoxycarbonyl group); B⁶ represents a hydroxyl-protecting group (preferably a benzyl group); and Z represents an optically active 2-oxo-oxazolidinyl group. The compound (4) is either known from Massall, C. M., Johnson M. and Theobald C. J. [J. Chem. Soc. Perkin I, (1971), 1451] or can be produced according to the procedure of Steps 1a, 1b and 2 described below.

(Step 1a)

This step involves hydrolyzing a compound (2), which is known from Synthetic Communications 1988, 2225, in an inert solvent in the presence of a base.

There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material to some extent. Examples of such solvents include: aliphatic hydrocarbons such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons such as benzene, toluene or xylene; halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, isoamyl alcohol, diethylene glycol, glycerin, octanol, cyclohexanol or methyl cellosolve; ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitro compounds such as nitroethane or nitrobenzene; nitriles such as acetonitrile or isobutyronitrile; amides such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides such as dimethyl sulfoxide or sulfolane; preferably alcohols.

There is no particular limitation upon the nature of the base used, provided that it can be used as a base in conventional reactions. Examples of such bases include: alkali metal carbonates such as sodium carbonate, potassium carbonate or lithium carbonate; alkali metal hydrogencarbonates such as sodium hydrogencarbonate, potassium hydrogencarbonate or lithium hydrogencarbonate; alkali metal hydrides such as lithium hydride, sodium hydride or potassium hydride; and alkali metal hydroxides such as sodium hydroxide, potassium hydroxide, barium hydroxide or lithium hydroxide; preferably alkali metal hydroxides.

The reaction is usually carried out at a temperature of 20° to 200° C., preferably 50° to 130° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 2 to 72 hours, preferably 5 to 24 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example, by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid such as hydrochloric acid or sulfuric acid; extracting with a water-immiscible solvent such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used for the following reaction without further purification but, if desired, can be purified by conventional means such as chromatography or recrystallization.

(Step 1b)

This step consists in protecting the amino group at the 1-position of the piperazic acid prepared in Step 1a in an inert solvent to produce a compound (3).

As a preferred amino-protecting group there come into consideration aralkyloxycarbonyl groups such as the benzyloxycarbonyl group.

The reaction can be carried out by using a reagent for preparing carbamates (preferably benzyloxycarbonyl chloride), which is commercially available or can readily be prepared, in an inert solvent in the presence of a base. There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effects upon the reaction and can dissolve the starting material to some extent. Examples of such solvents include: aliphatic hydrocarbons such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons such as benzene, toluene or xylene; alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, isoamyl alcohol, diethylene glycol, glycerin, octanol, cyclohexanol or methyl cellosolve; water; and a mixture of water and one or more of these organic solvents; preferably a mixture of water and the corresponding alcohols.

There is no particular limitation upon the nature of the base used, provided that it can be used as a base in conventional reactions. Examples of such bases include: inorganic bases, including alkali metal carbonates such as sodium carbonate, potassium carbonate or lithium carbonate; alkali metal hydrogencarbonates such as sodium hydrogencarbonate, potassium hydrogencarbonate or lithium hydrogencarbonate; alkali metal hydrides such as lithium hydride, sodium hydride or potassium hydride; and alkali metal hydroxides such as sodium hydroxide, potassium hydroxide, barium hydroxide or lithium hydroxide; and organic bases such as triethylamine, tributylamine, diisopropylethylamine, N-methylmorpholine, pyridine, 4-(N,N-dimethylamino)pyridine, N,N-dimethylaniline, N,N-diethylanilene, 1,5-diazabicyclo[4,3,0]non-5-ene (DBN), 1,4-diazabicyclo[2,2,2]octane (DABCO) or 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU); preferably alkali metal hydroxides or organic bases.

The reaction is usually carried out at a temperature of 0° to 50° C., preferably 0° to 20° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 24 hours, preferably 1 to 3 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example, by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid such as Hydrochloric acid or sulfuric acid; extracting with a water-immiscible solvent such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used in the following reaction without further purification but, if desired, can be purified by conventional means such as chromatography or recrystallization.

(Step 2)

In this step, a compound (4) can be prepared 1 by reacting a compound (3) with an alcohol, B¹ OH, (particularly benzyl alcohol or tert-butyl alcohol) in an inert solvent in the presence of a condensing agent or 2 by reacting a compound (3) with an esterifying agent in an inert solvent.

In process 1, the nature of the solvent used is not particularly critical, provided that it has no adverse effect upon the reaction and can dissolve the starting material to some extent. Examples of preferred solvents include: aliphatic hydrocarbons such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons such as benzene, toluene or xylene; halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl acetate, butyl acetate or diethyl carbonate; ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitro compounds such as nitroethane or nitrobenzene; nitriles such as acetonitrile or isobutyronitrile; amides such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides such as dimethyl sulfoxide or sulfolane; preferably aromatic hydrocarbons, ethers, halogenated hydrocarbons, nitriles or amides.

Examples of the condensing agents used include: for example, di(lower alkyl) azodicarboxylate-triphenylphosphine, such as diethyl azodicarbonylate-triphenylphosphine; N-(lower alkyl)-5-arylisoxazolium-3'-sulfonates such as N-ethyl-5-phenylisoxazolium-3'-sulfonate; N,N'-dicycloalkylcarbodiimides such as N,N'-dicyclohexylcarbodiimide (DCC); diheteroaryl diselenides such as dipyridyl diselenide; phosphines such as diethyl phosphoryl cyanide (DEPC); arylsulfonyl triazolides such as p-nitrobenzenesulfonyl triazolide; 2-halo-1-(lower alkyl)pyridinium halides such as 2-chloro-1-methylpyridinium iodide; diarylphosphoryl azides such as diphenylphosphoryl azide; imidazole derivatives such as N,N'-carbodiimidazole (CDI); benzotriazole derivatives such as 1-hydroxybenzotriazole (HOBT); dicarboximide derivatives such as N-hydroxy-5-norbornene-2,3-dicarboximide (HONB); and carbodiimides such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAPC); preferably carbodiimides (particularly DCC and DEPC).

The reaction is usually carried out at a temperature of 0° to 100° C., preferably 10° to 50° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 48 hours, preferably 1 to 12 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example, by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid such as hydrochloric acid or sulfuric acid; filtering off insoluble materials; extracting with a water-immiscible solvent such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used in the following reaction without further purification but if desired, can be purified by conventional means such as chromatography or recrystallization.

In the process 2, the desired compound can be prepared by reacting with an esterifying agent, preferably isobutene, in an inert solvent in the presence of an acid.

Examples of the solvents used include: aliphatic hydrocarbons such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons such as benzene, toluene or xylene; halogenated hydrocarbons such as dichloromethane, chlorobenzene or dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl acetate, butyl acetate or diethyl carbonate; ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitro compounds such as nitroethane or nitrobenzene; nitriles such as acetonitrile or isobutyronitrile; amides such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides such as dimethyl sulfoxide or sulfolane; preferably ethers (particularly dioxane).

There is no particular limitation upon the nature of the acid catalyst used, provided that it can be used as an acid catalyst in conventional reactions. Preferred examples of such acid catalysts include; Bronsted acids including inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, perchloric acid or phosphoric acid; and organic acids such as acetic acid, formic acid, oxalic acid, methanesulfonic acid, p-toluenesulfonic acid, trifluoroacetic acid or trifluoromethanesulfonic acid; or Lewis acids such as zinc chloride, tin tetrachloride, boron trichloride, boron trifluoride or boron tribromide, preferably inorganic acids; and more preferably strong inorganic acids (particularly hydrochloric acid).

The reaction is usually carried out at a temperature of 0° to 50° C., preferably 0° to 25° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 48 hours, preferably 1 to 24 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example, by distilling off the solvent; pouring the reaction mixture into water; extracting it with a water-immiscible solvent such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used in the following reaction without further purification but if desired, can be purified by conventional means such as chromatography or recrystallization.

(Step 3 )

In this step, a compound (6) can be prepared 1 by reacting a activated derivative (that is, an acid halide or acid anhydride) of a malonic acid (5) with the corresponding alcohol, B² OH, (particularly methanol or ethanol) in the presence of a base or 2 by reacting a malonic acid (5) with the corresponding alcohol, B² OH, in the presence of a condensing agent.

In the process 1, there is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of such solvents include: aliphatic hydrocarbons such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons such as benzene, toluene or xylene; halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl acetate, butyl acetate or diethyl carbonate; ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitro compounds such as nitroethane or nitrobenzene; nitriles such as acetonitrile or isobutyronitrile; amides such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides such as dimethyl sulfoxide or sulfolane; preferably alcohols, aromatic hydrocarbons or halogenated hydrocarbons.

As a halide moiety of the acid halide used there come into consideration chlorine, bromine and iodine, preferably chlorine or bromine.

The nature of the base used in the reaction is not particularly critical, provided that it can be used as a base in conventional reactions. Examples of such bases include: inorganic base salts including alkali metal carbonates such as sodium carbonate, potassium carbonate or lithium carbonate; alkali metal hydrogencarbonates such as sodium hydrogencarbonate, potassium hydrogencarbonate or lithium hydrogencarbonate; alkali metal hydrides such as lithium hydride, sodium hydride or potassium hydride; and alkali metal hydroxides such as sodium hydroxide, potassium hydroxide, barium hydroxide or lithium hydroxide; or organic bases such as triethylamine, tributylamine, diisopropylethylamine, N-methylmorpholine, pyridine, 4-(N,N-dimethylamino)pyridine, N,N-dimethylaniline, N,N-diethylaniline, 1,5-diazabicyclo[4,3,0]non-5-ene (DBN), 1.4-diazabicyclo[2,2,2]octane (DABCO) or 1,8-diazabicyclo-[5,4,0]undec:7-ene (DBU); preferably alkali metal hydroxides or organic bases.

The reaction is usually carried out at a temperature of 0° to 60° C., preferably 0° to 30° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 24 hours, preferably 1 to 6 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example, by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid such as hydrochloric acid or sulfuric acid; extracting it with a water-immiscible solvent such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can be used in the following reaction without further purification but, if desired, can be purified by conventional means such as chromatography or recrystallization.

In the process 2, the nature of the solvent used is not particularly critical, providing that it has no adverse effect upon the reaction and can dissolve the starting material to some extent. Examples of such solvents include: aliphatic hydrocarbons such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons such as benzene, toluene or xylene; halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl acetate, butyl acetate or diethyl carbonate; ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitro compounds such as nitroethane or nitrobenzene; nitriles such as acetonitrile or isobutyronitrile; and amides such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; preferably halogenated hydrocarbons (particularly dichloromethane), ethers (particularly tetrahydrofuran) or aromatic hydrocarbons (particularly benzene).

Preferred examples of the condensing agents used include: DCC (dicyclohexylcarbodiimide), CDI (N,N'-carbonyldiimidaole), DPPA (diphenylphosphoryl azide), HOBT (1-hydroxybenzotriazole), HONB (N-hydroxy-5-norbornene-2,3-dicarboximide) and EDAPC [1-ethyl-3-(3-dimethylaminopropyl)carbodiimide].

In the case where a condensing agent is used, the reaction is more efficiently effected by carrying it out in combination with a deacidifying agent.

Preferred examples of the deacidifying agent used include: organic amines, such as pyridine, dimethylaminopyridine or pyrrolidinopyridine. In the case where the said condensing agent is used, the reaction is accelerated by using these amines.

The reaction is usually carried out at a temperature of 0° to 60° C., preferably 0° to 30° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 48 hours, preferably 1 to 12 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, separated and purified by various means in a proper combination. An example of one such technique comprises: pouring the reaction mixture into water; adding a water-immiscible solvent such as benzene, ether or ethyl acetate; filtering off insoluble materials, if any; separating an organic solvent layer; washing the extract with diluted hydrochloric acid or an aqueous solution of sodium hydrogencarbonate; and finally distilling off the solvent. The desired product thus obtained, if necessary, can be purified by conventional means, such as adsorption or ion exchange chromatography through various carriers, such as activated charcoal or silica gel; gel filtration through sephadex; or recrystallization from an organic solvent such as ether, ethyl acetate or chloroform.

(Step 4)

In this step, a compound (7) can be prepared by reacting a compound (6) with an alkyl halide R² X (wherein X signifies a halogen atom, preferably chlorine or bromine), in an inert solvent in the presence of base.

There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of such solvents include: aliphatic hydrocarbons, such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons, such as benzene, toluene or xylene; halogenated hydrocarbons, such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; esters, such as ethyl formate, ethyl acetate, propyl acetate, butyl acetate or diethyl carbonate; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; alcohols, such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, isoamyl alcohol, diethylene glycol, glycerin, octanol, cyclohexanol or methyl cellosolve; ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitro compounds, such as nitroethane or nitrobenzene; nitriles, such as acetonitrile or isobutyronitrile; amides, such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides, such as dimethyl sulfoxide or sulfolane; preferably ethers (particularly diethyl ether or tetrahydrofuran), amides (particularly dimethylformamide).

The nature of the base used is not particularly critical, provided that it can be used as a base in conventional reactions. Examples of the bases include: inorganic base salts, including alkali metal carbonates, such as sodium carbonate, potassium carbonate or lithium carbonate; alkali metal hydrogencarbonates, such as sodium hydrogencarbonate, potassium hydrogencarbonate or lithium hydrogencarbonate; alkali metal hydrides, such as lithium hydride, sodium hydride or potassium hydride; alkali metal alkoxides; such as sodium methoxide, sodium ethoxide or potassium butoxide; and alkali metal hydroxides, such as sodium hydroxide, potassium hydroxide, barium hydroxide or lithium hydroxide; or organic bases, such as triethylamine, tributylamine, diisopropylethylamine, N-methylmorpholine, pyridine, 4-(N,N-dimethylamino)pyridine, N,N-dimethylaniline, N,N-diethylaniline, 1,5-diazabicyclo[4,3,0]non-5-ene (DBN), 1.4-diazabicyclo[2,2,2]octane (DABCO) or 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU); preferably alkali metal hydroxides, alkali metal alkoxides or organic bases.

As a halide moiety of the alkyl halide used there come into consideration chlorine, bromine and iodine, preferably chlorine or bromine.

The reaction is usually carried out at a temperature of -20° to 100° C., preferably -10° to 50° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 48 hours, preferably 1 to 15 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, separated and purified by various means in a proper combination. An example of one such technique comprises: pouring the reaction mixture into water; adding a water-immiscible solvent, such as benzene, ether or ethyl acetate; filtering off insoluble materials, if any; separating an organic solvent layer; washing the extract with diluted hydrochloric acid or an aqueous solution of sodium hydrogencarbonate; and finally distilling off the solvent. The desired product thus obtained, if necessary, can be purified by conventional means, such as adsorption or ion exchange chromatography through various carriers, such as activated charcoal or silica gel; gel filtration through sephadex; or recrystallization from an organic solvent, such as ether, ethyl acetate or chloroform.

(Step 5)

In this step, a compound (8) can be prepared by subjecting a compound (7) to a hydrolysis reaction with a base in an inert solvent, followed by decarboxylation.

Examples of the solvents used hydrocarbons, such hydrocarbons, such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons, such as benzene, toluene or xylene; halogenated hydrocarbons, such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; alcohols, such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, isoamyl alcohol, diethylene glycol, glycerin, octanol, cyclohexanol or methyl cellosolve; ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitro compounds, such as nitroethane or nitrobenzene; nitriles, such as acetonitrile or isobutyronitrile; amides, such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; sulfoxides, such as dimethyl sulfoxide or sulfolane; and a mixture of water and one or more of these organic solvents; preferably a mixture of alcohols and water.

The nature of the base used is not particularly critical, provided that it can be used as a base in conventional reactions. Examples of the bases used include: inorganic base salts, including alkali metal carbonates, such as sodium carbonate, potassium carbonate or lithium carbonate; alkali metal hydrogencarbonates, such as sodium hydrogencarbonate, potassium hydrogencarbonate or lithium hydrogencarbonate; alkali metal hydrides, such as lithium hydride, sodium hydride or potassium hydride; and alkali metal hydroxides, such as sodium hydroxide, potassium hydroxide, barium hydroxide or lithium hydroxide; or organic bases, such as triethylamine, tributylamine, diisopropylethylamine, N-methylmorpholine, pyridine, 4-(N,N-dimethylamino)pyridine, N,N-dimethylaniline, N,N-diethylaniline, 1,5-diazabicyclo[4,3,0]non-5-ene (DBN), 1,4-diazabicyclo[2,2,2]octane (DABCO) or 1.8-diazabicyclo[5,4,0]undec-7-ene (DBU); preferably alkali metal hydroxides or organic bases.

The reaction is usually carried out at a temperature of 0° to 120° C., preferably 20° to 120° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 48 hours, preferably 1 to 16 hours.

In this step, the hydrolysis of a malonate derivative is generally accompanied by decarboxylation. However, where only the hydrolysis proceeds and decarboxylation is incomplete, the reaction can be accomplished by adding an organic base, such as collidine or lutidine, followed by heating at 100° to 120° C.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, separated and purified by various means in a proper combination. An example of one such technique comprises: pouring the reaction mixture into water; acidifying with hydrochloric acid etc.; adding a water-immiscible solvent, such as benzene, ether or ethyl acetate; filtering off insoluble materials, if any; separating an organic solvent layer; washing the extract with diluted hydrochloric acid; and, finally, distilling off the solvent. The desired product thus obtained, if necessary, can be purified by conventional means, such as: adsorption or ion exchange chromatography through various carriers, such as activated charcoal or silica gel; gel filtration through a sephadex column; or recrystallization from an organic solvent, such as ether, ethyl acetate or chloroform.

(Step 6)

In this step, a compound (9) can be prepared from a compound (8), which is prepared in Step 5 or which is commercially available, and the reaction is conducted in a similar manner to that of Step 3.

(Step 7)

In this step, a compound (10) can be prepared by reacting an acid halide, which is prepared by reacting a compound (8) with a reagent for the halogenation of a carboxylic acid, with an optically active oxazolidinone ZH in an inert solvent in the presence of n-butyllithium.

The nature of the solvent used in the halogenation is not critical, providing that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of preferred solvents include: aliphatic hydrocarbons, such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons, such as benzene, toluene or xylene; and halogenated hydrocarbons, such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene.

Examples of the reagents used for halogenation include: thionyl halides, such as thionyl chloride or thionyl bromide; and phosphorus compounds, such as phosphorus trichloride, phosphorus tribromide, phosphorus pentachloride or phosphorus oxychloride.

The reaction is usually carried out at a temperature of 0° to 50° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 0.2 to 5 hours, preferably 0.2 to 1 hour.

After completion of the reaction, the desired compound can be isolated from the reaction mixture by distilling off the solvent. The product thus obtained can be used in the following reaction without further purification.

The nature of the solvent used in the reaction with a 2-oxazolidinone derivative is not particularly critical, providing that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of particularly preferred solvents include: ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether.

Examples of optically active 2-oxazolidinone derivatives include: (4S)-isopropyl-2-oxazolidinone, (4R)-isopropyl-2-oxazolidinone, (4S)-benzyl-2-oxazolidinone and (4R)-benzyl-2-oxazolidinone.

The reaction is usually carried out at a temperature of -78° to 20° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 0.2 to 5 hours, preferably 0.2 to 2 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example: by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid, such as hydrochloric acid or sulfuric acid; extracting with a water-immiscible solvent, such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used in the following reaction without further purification but if desired, can be purified by conventional means, such as chromatography or recrystallization.

(Step 8)

In this step, a compound (11) can be prepared by reacting a compound (9) with a tri(substituted) silyl halide, B³ X (wherein X is as defined above), in an inert solvent in the presence of a base.

There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of such solvents include: aliphatic hydrocarbons, such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons, such as benzene, toluene or xylene; halogenated hydrocarbons, such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; and nitriles, such as acetonitrile or isobutyronitrile.

The nature of the base used is not particularly critical, provided that it can be used as a base in conventional reactions. Examples of preferred bases include: alkali metal carbonates, such as sodium carbonate, potassium carbonate or lithium carbonate; alkali metal hydrogencarbonates, such as sodium hydrogencarbonate, potassium hydrogencarbonate or lithium hydrogencarbonate; alkali metal hydrides, such as lithium hydride, sodium hydride or potassium hydride; organic bases, such as triethylamine, tributylamine, diisopropylethylamine, N-methylmorpholine, pyridine, 4-(N,N-dimethylamino)pyridine, N,N-dimethylaniline, N,N-diethylaniline, 1,5-diazabicyclo[4,3,0]non-5-ene (DBN), 1,4-diazabicyclo[2,2,2]octane (DABCO) or 1,8-diazabicyclo[5,4,0 ]undec-7-ene (DBU); and organic metal bases, such as butyllithium or lithium diisopropylamide.

Examples of the trialkylsilyl halides used include: trimethylsilyl chloride or tert-butyldimethylsilyl chloride, preferably trimethylsilyl chloride.

The reaction is usually carried out at a temperature of -78° to 20° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 10 hours.

After completion of the reaction, the desired compound can be recovered from the reaction mixture, for example: by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid, such as hydrochloric acid or sulfuric aicd; extracting with a water-immiscible solvent, such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used in the following reaction without further purification but, if desired, can be purified by conventional means, such as chromatography or recrystallization.

(Step 9)

In this step, a compound (12) can be prepared by reacting a compound (10) with an α-haloacetate XCH₂ -COOB⁴ (wherein X is as defined above), in an inert solvent in the presence of a base.

There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of preferred solvents include: aliphatic hydrocarbons, such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons, such as benzene, toluene or xylene; halogenated hydrocarbons, such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; alcohols, such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, isoamyl alcohol, diethylene glycol, glycerin, octanol, cyclohexanol or methyl cellosolve; amides, such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides, such as dimethyl sulfoxide or sulfolane; preferably ethers.

The nature of the base used is not particularly critical, provided that it can be used as a base in conventional reactions. Preferred examples of preferred bases include: alkali metal carbonates, such as sodium carbonate, potassium carbonate or lithium carbonate; alkali metal hydrogencarbonates, such as sodium hydrogencarbonate, potassium hydrogencarbonate or lithium hydrogencarbonate; alkali metal hydrides, such as lithium hydride, sodium hydride or potassium hydride; alkali metal alkoxides, such as sodium methoxide, sodium ethoxide, potassium tert-butoxide or lithium methoxide; alkali metal mercaptides, such as sodium methylmercaptide or sodium ethylmercaptide; organic bases, such as triethylamine, tributylamine, diisopropylethylamine, N-methylmorpholine, pyridine, 4-(N,N-dimethylamino)pyridine, N,N-dimethylaniline, N,N-diethylaniline, 1,5-diazabicyclo[4,3,0]non-5-ene (DBN), 1,4-diazabicyclo[2,2,2]octane (DABCO) or 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU); and organic metal bases, such as butyllithium or lithium diisopropylamide; preferably lithium diisopropylamide (in tetrahydrofuran).

As a halo moiety of the α-haloacetate used there come into consideration chlorine, bromine and iodine.

As an ester moiety of the α-haloacetate used there come into consideration methyl, ethyl, benzyl and tert-butyl.

The reaction is usually carried out at a temperature of -78° to 10° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 10 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example: by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid, such as hydrochloric acid or sulfuric acid; extracting with a water-immiscible solvent, such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used in the following reaction without further purification but, if desired, can be purified by conventional means, such as chromatography or recrystallization.

(Step 10)

In this step, a compound (13) can be prepared by hydrolysis of a compound (12) in an inert solvent in the presence of a base to remove an optically active 2-oxazolidinone derivative.

There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of preferred solvents include: aliphatic hydrocarbons, such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons, such as benzene, toluene or xylene; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane, diethylene glycol or diethylene glycol dimethyl ether; alcohols, such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, isoamyl alcohol, diethylene glycol, glycerin, octanol, cyclohexanol or methyl cellosolve; ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitro compounds, such as nitroethane or nitrobenzene; nitriles, such as acetonitrile or isobutyronitrile; amides, such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides, such as dimethyl sulfoxide or sulfolane.

The nature of the base used is not particularly critical, provided that it can be used as a base in conventional reactions. Examples of preferred bases include: alkali metal carbonates, such as sodium carbonate, potassium carbonate or lithium carbonate; alkali metal hydrogencarbonates, such as sodium hydrogencarbonate, potassium hydrogencarbonate or lithium hydrogencarbonate; alkali metal hydrides, such as lithium hydride, sodium hydride or potassium hydride: alkali metal hydroxides, such as sodium hydroxide, potassium hydroxide, barium hydroxide or lithium hydroxide; alkali metal alkoxides, such as sodium methoxide, sodium ethoxide, potassium tert-butoxide or lithium methoxide; alkali metal mercaptides, such as sodium methylmercaptide or sodium ethylmercaptide; and organic bases, such as triethylamine, tributylamine, diisopropylethylamine, N-methylmorpholine, pyridine, 4-(N,N-dimethylamino)pyridine, N,N-dimethylaniline, N,N-diethylaniline, 1,5-diazabicyclo[4,3,0]non-5-ene (DBN), 1,4-diazabicyclo[2,2,2]octane (DABCO) or 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU).

In this step, it is most preferable that the reaction is conducted using lithium hydroxide in a mixture of tetrahydrofuran and water in the presence of hydrogen peroxide.

The reaction is usually carried out at a temperature of 0° to 100° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 24 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example: by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid, such as hydrochloric acid or sulfuric acid; extracting with a water-immiscible solvent, such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used in the following reaction without further purification but, if desired, can be purified by conventional means, such as chromatography or recrystallization.

(Step 11)

In this step, a compound (14) can be prepared by reacting a compound (11) with an α-halocarboxylate XCH₂ -COOB⁴ (wherein X is as defined above), in an inert solvent.

There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of preferred solvents include: aliphatic hydrocarbons, such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons, such as benzene, toluene or xylene; halogenated hydrocarbons, such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; esters, such as ethyl formate, ethyl acetate, propyl acetate, butyl acetate or diethyl carbonate; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; nitriles, such as acetonitrile or isobutyronitrile; amides, such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides, such as dimethyl sulfoxide or sulfolane.

As a halo moiety of the α-halocarboxylate used in the reaction there come into consideration chlorine, bromine and iodine, preferably chlorine or bromine. As an ester moiety of the α-halocarboxylate used in the reaction there come into consideration methyl, ethyl, tert-butyl and benzyl.

The reaction is usually carried out at a temperature of -78° to 30° C. The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 0.5 to 10 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example: by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid, such as hydrochloric acid or sulfuric acid; extracting with a water-immiscible solvent, such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used in the following reaction without further purification but if desired, can be purified by conventional means, such as chromatography or recrystallization.

(Step 12)

In this step, a compound (13) can be prepared by hydrolyzing a compound (14) in an inert solvent in the presence of a base.

There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of preferred solvents include: aliphatic hydrocarbons, such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons, such as benzene, toluene or xylene; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; alcohols, such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, isoamyl alcohol, diethylene glycol, glycerin, octanol, cyclohexanol or methyl cellosolve; ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitriles, such as acetonitrile or isobutyronitrile; amides, such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides, such as dimethyl sulfoxide or sulfolane.

The nature of the base used in the reaction is not particularly critical, provided that it can be used as a base in conventional reactions. Examples of preferred bases include: alkali metal carbonates, such as sodium carbonate, potassium carbonate or lithium carbonate; alkali metal hydrogencarbonates, such as sodium hydrogencarbonate, potassium hydrogencarbonate or lithium hydrogencarbonate; alkali metal hydrides, such as lithium hydride, sodium hydride or potassium hydride; alkali metal hydroxides, such as sodium hydroxide, potassium hydroxide, barium hydroxide or lithium hydroxide; alkali metal alkoxides, such as sodium methoxide, sodium ethoxide, potassium tert-butoxide or lithium methoxide.

The reaction is usually carried out at a temperature of 0° to 100° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 24 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example: by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid, such as hydrochloric acid or sulfuric acid; extracting with a water-immiscible solvent, such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used in the following reaction without further purification but if desired, can be purified by conventional means, such as chromatography or recrystallization.

If desired, a protecting group, B⁴, of a compound (13) can be replaced by another protecting group. This exchange reaction is conducted by conventional transesterification or, after protecting a free carboxylic acid group in the molecule, the protecting group, B⁴, is eliminated according to the procedure described in Step 20, followed by introducing the desired protecting group and then reproducing a carboxylic acid group.

(Step 13)

In this step, a compound (18) can be prepared by reacting a compound (15), which is obtained by protecting an amino group of a commercially available α-amino acid by conventional means, with an alcohol, R⁸ OH, in an inert solvent in the presence of a condensing agent. The reaction is carried out in a manner similar to that of Step 2.

(Step 14)

In this step, a compound (16) can be prepared by reacting a compound (15) with an amine, R⁹ R¹⁰ NH, in an inert solvent in the presence of a condensing agent.

There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of preferred solvents include: aliphatic hydrocarbons, such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons, such as benzene, toluene or xylene; halogenated hydrocarbons, such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; esters; such as ethyl formate, ethyl acetate, propyl acetate, butyl acetate or diethyl carbonate; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitro compound, such as nitroethane or nitrobenzene; nitriles, such as acetonitrile or isobutyronitrile; amides, such as formamide, dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides, such as dimethyl sulfoxide or sulfolane.

Examples of suitable condensing agents include: di(lower alkyl) azodicarboxylate-triphenylphosphines such as diethyl azodicarboxylate-triphenylphosphine; N-(lower alkyl)-5-arylisoxazolium-3'-sulfonates, such as N-ethyl-5-phenylisoxazolium-3'-sulfonate; N,N'-dicycloalkylcarbodiimides, such as N,N'-dicyclohexylcarbodiimide (DCC); diheteroaryl diselenides, such as 2,2'-dipyridyl diselenide; phosphins, such as diethylphosphoryl cyanide (DEPC); arylsulfonyl triazolides, such as p-nitrobenzenesulfonyl triazolide; 2-halo-1-(lower alkyl)pyridinium halides, such as 2-chloro-1-methylpyridinium iodide; diarylphosphoryl azides, such as diphenylphosphoryl azide (DPPA); imidazole derivatives, such as N,N'-carbodiimidazole (CDI); benzotriazole derivatives, such as 1-hydroxybenzotriazole (HOBT); dicarboximide derivatives, such as N-hydroxy-5-norbornene-2,3-dicarboximide (HONB), and carbodiimide derivatives, such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAPC); preferably diarylphosphoryl azide.

The reaction is usually carried out at a temperature of 0° to 150° C., preferably 20° to 100° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 100 hours, preferably 2 to 24 hours.

After completion of the reaction, the desired compound can be isolated from the reaction mixture, for example: by neutralizing the reaction mixture properly; filtering off insoluble materials, if any; adding water and a water-immiscible organic solvent, such as ethyl acetate, separating an organic layer; washing the extract with water; and distilling off the solvent from the extract after drying over a drying agent, such as anhydrous magnesium sulfate.

(Step 15)

In this step, a compound (17) can be prepared by reacting a compound (16) with a reagent for the deprotection of an amino-protecting group in an inert solvent.

The procedure used for the deprotection depends upon the nature of the amino-protecting group but the reaction is carried out as follows.

Where an amino-protecting group is a silyl group, it can be eliminated by treating with a compound capable of forming a fluorine anion, such as tetrabutylammonium fluoride.

The reaction temperature and the time required for the reaction are not particularly critical, but the reaction is usually conducted at ambient temperature for a period of 10 to 18 hours.

Where an amino-protecting group is an alkoxycarbonyl group or a substituted methylene group capable of forming a Schiff's base, it can be eliminated by treating with an acid in a water soluble solvent.

There is no particular limitation upon the nature of the acid used, provided that it can usually be used as an acid in conventional reactions and has no adverse effect upon the reaction. Examples of preferred acids include inorganic acids, such as hydrochloric acid, sulfuric acid, phosphoric acid or hydrobromic acid.

There is no particular limitation upon the nature of the solvent used, provided that it can be used in conventional hydrolysis. Examples of preferred solvents include: water; organic solvents including alcohols, such as methanol, ethanol or n-propanol; and ethers, such as tetrahydrofuran or dioxane; or a mixture of water and one or more these organic solvents.

The reaction temperature and the time required for the reaction vary depending upon the nature of the starting material and of the solvent as well as the nature of the acid or base used. Although there is no particular limitation, the reaction is usually carried out at a temperature of 0° to 150° C. for a period of 1 to 10 hours in order to suppress side reactions.

Where an amino-protecting group is an aralkyl or aralkyloxycarbonyl group, it can preferably be removed by contacting with a reducing agent (preferably catalytic reduction at ambient temperature in the presence of a catalyst) or with an oxidizing agent in a solvent.

In the case of deprotection by catalytic reduction, there is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction. Examples of preferred solvents include: alcohols, such as methanol, ethanol or isopropanol; ethers, such as diethyl ether, tetrahydrofuran or dioxane; aromatic hydrocarbons, such as benzene, toluene or xylene; aliphatic hydrocarbons, such as hexane or cyclohexane; esters, such as ethyl acetate or propyl acetate; fatty acids, such as acetic acid; and a mixture of one or more of these organic solvents and water.

There is no particular limitation upon the nature of the catalyst used, provided that it can be used as a catalyst in conventional catalytic reductions. Examples of preferred catalysts include: palladium on charcoal, Raney nickel, platinum oxide, platinum black, rhodium on alumina, triphenylphosphine-rhodium chloride and palladium on barium sulfate.

There is no particular limitation upon the pressure used and the reaction is usually carried out at a pressure of 1 to 10 atmospheric pressures.

The reaction temperature and the time required for the reaction vary depending upon the nature of the starting material and of the solvent but the reaction is usually carried out at a temperature of 0° to 100° C. for a period of 5 minutes to 24 hours.

In the case of deprotection by oxidation, the solvent used is not particularly critical, provided that it has no adverse effect upon the reaction. A preferred solvent is an aqueous organic solvent.

Preferred examples of such solvents include: ketones, such as acetone; halogenated hydrocarbons, such as dichloromethane, chloroform or carbon tetrachloride; nitriles, such as acetonitrile; ethers, such as diethyl ether, tetrahydrofuran or dioxane; amides, such as dimethylformamide, dimethylacetamide or hexamethylphosphoramide; and sulfoxides, such as dimethyl sulfoxide.

There is no particular limitation upon the nature of the oxidizing agent used, provided that it can be used in conventional oxidation. Examples of preferred oxidizing agents include: potassium persulfate, sodium persulfate, ammonium cerium nitrate (CAN) and 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ).

Although the reaction temperature and the time required for the reaction vary depending upon the nature of the starting material and of the solvent as well as the type of the catalyst used, the reaction is usually conducted at a temperature of 0° to 150° C. for a period of 10 minutes to 24 hours.

Where an amino-protecting group is an alkenyloxycarbonyl group, it can usually be removed by treating with a base in a similar manner as where an amino-protecting group is the said aliphatic acyl, aromatic aryl or alkoxycarbonyl group, or a substituted methylene group capable of forming a Schiff's base.

Where an amino-protecting group is an aryloxycarbonyl group, it can simply be eliminated by using palladium, triphenylphosphine or nickel tetracarbonyl and the side reactions are suppressed.

The reaction temperature depends upon the nature of the starting material and of the solvent as well as the type of the reagent to be used, but the reaction is usually carried out at a temperature of 0° to 150° C., preferably 20° to 100° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is usually complete within a period of 1 to 100 hours, preferably 2 to 24 hours.

After completion of the reaction, the desired compound is isolated from the reaction mixture, for example: by neutralizing the reaction mixture properly; filtering off insoluble materials, if any; adding water and a water-immiscible organic solvent, such as ethyl acetate; separating the organic layer; washing the extract with water; drying over a drying agent, such as anhydrous magnesium sulfate; and distilling off the solvent. The desired product thus obtained can be purified, if necessary, by conventional means, such as recrystallization, reprecipitation or chromatography.

(Step 16)

In this step, a compound (19) is prepared by reacting a compound (18) with a reagent for the deprotection of an amino-protecting group in an inert solvent, and the reaction is conducted in a manner similar to that of Step 15.

(Step 17)

In this step, a compound (20) can be prepared by reacting a compound (16) (provided that either of R⁹ or R¹⁰ is a methoxy group when the other group is methyl) with a reagent R⁷ M [wherein M signifies alkali metals, such as lithium, or a Grignard reagent (MgBr or MgCl)] in an inert solvent.

There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of such solvents include: aliphatic hydrocarbons, such as hexane, heptane, ligroin or petroleum ether; aromatic hydrocarbons, such as benzene, toluene or xylene; halogenated hydrocarbons, such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether.

The reaction temperature depends upon the nature of the starting material and of the solvent as well as the type of the reagent used, but the reaction is usually carried out at a temperature of -70° to 50° C., preferably -40° to 30° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent, but the reaction is complete within a period of 5 minutes to 24 hours, preferably 30 minutes to 3 hours.

After completion of the reaction, the desired compound is isolated from the reaction mixture, for example: by neutralizing the reaction mixture properly; filtering off insoluble materials, if any; adding water and a water-immiscible organic solvent, such as ethyl acetate; separating the organic layer; washing the extract with water; drying over a drying agent, such as anhydrous magnesium sulfate; and distilling off the solvent.

The desired product thus obtained can be purified, if necessary, by conventional means, such as recrystallization, reprecipitation or chromatography.

(Step 18)

In this step, a compound (21) can be prepared by reacting a compound (20) with a reagent for the deprotection of an amino-protecting group in an inert solvent, and the reaction is conducted in a manner similar to that of Step 15.

(Step 19)

In this step, a compound (22) can be prepared by reacting a compound (4) with a compound (13) in an inert solvent in the presence of a condensing agent, and the reaction is conducted in a manner similar to that of Step 14.

(Step 20)

In this step, a compound (23) can be prepared by removing a protecting group, B⁴, of a compound (22) in an inert solvent.

Deprotecting procedures depend upon the nature of the protecting group used, but can be conducted according to a method well-known in this art field.

Where a carboxyl-protecting group is a tert-butyl or benzhydryl group, it is deprotected by treating with trifluoroacetic acid, hydrobromic acid in acetic acid or hydrochloric acid in dioxane.

Where a carboxyl-protecting group is a trichloroethyl or trichlorobromo group, it is deprotected by reacting with zinc powder in a mixture of acetic acid or a phosphate buffer solution (pH=4.27-7.2) and ethers, such as tetrahydrofuran.

Where a carboxyl-protecting group is an aralkyl group, such as a benzyl group, it is deprotected by contacting with a reducing agent in a solvent (preferably catalytic reduction at ambient temperature in the presence of a catalyst).

(Step 21)

In this step, a compound (24) can be prepared by reacting a compound (23) with a hydroxylamine, B⁶ ONH₂, in an inert solvent and the reaction is conducted in a manner similar to that of Step 19.

(Step 22)

In this step, a compound (1A) can be prepared by removing the protecting groups, A and B⁶ of a compound (24) in an inert solvent, with the proviso that B¹ has the same significance as R³.

Miscellaneous methods are conducted in this step depending upon the nature of the protecting groups.

For example, in the case of a compound (24) wherein B⁶ represents a benzyl group and A represents a benzyloxycarbonyl group, such protecting groups can be removed by catalytic reduction, which is carried out in a suitable solvent, such as alcohol, ether or acetic acid in the presence of a catalyst, such as palladium on charcoal or platinum in a stream of hydrogen. Where both A and B⁶ represent a tert-butoxycarbonyl group, elimination of a protecting group can be accomplished under an acidic condition using trifluoroacetic acid, hydrobromic acid in acetic acid or hydrochloric acid in dioxane. In all cases, the deprotection can be accomplished under the commonly used conditions for the deprotection of an amino- or hydroxyl-protecting groups.

There is no particular limitation upon the nature of the solvent used, provided that it has no adverse effect upon the reaction and can dissolve the starting material in some extent. Examples of such solvents include: aliphatic hydrocarbons, such as hexane, heptane, ligroin or petroleum ether, aromatic hydrocarbons, such as benzene, toluene or xylene; halogenated hydrocarbons, such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene or dichlorobenzene, esters, such as ethyl formate, ethyl acetate, propyl acetate, butyl acetate or diethyl carbonate; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane or diethylene glycol dimethyl ether; alcohols, such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, isoamyl alcohol, diethylene glycol, glycerin, octanol, cyclohexanol or methyl cellosolve; ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone or cyclohexanone; nitro compounds, such as nitroethane or nitrobenzene; nitriles, such as acetonitrile or isobutyronitrile; amides, such as formamide, dimethylformamide or hexamethylphosphoramide; sulfoxides, such as dimethyl sulfoxide or sulfolane.

The reaction is carried out at a temperature of -20° to 100° C., preferably 0° to 40° C.

The time required for the reaction depends upon the reaction temperature and other factors, such as the nature of the starting material and of the solvent used, but the reaction is usually complete within a period of 0.5 to 48 hours, preferably 1 to 5 hours.

After completion of the reaction, the desired compound is isolated from the reaction mixture, for example: by distilling off the solvent; pouring the reaction mixture into water; acidifying with an inorganic acid, such as hydrochloric acid or sulfuric acid; extracting with a water-immiscible solvent, such as benzene, ether or ethyl acetate; and distilling off the solvent from the extract. The product thus obtained can usually be used in the following reaction without further purification but if desired, can be purified by conventional means, such as chromatography or recrystallization.

(Step 23)

In this step, a compound (25) can be prepared by eliminating a protecting group, B¹ of a compound (24) in an inert solvent and the reaction is conducted in a manner similar to that of Step 20.

(Step 24)

In this step, a compound (26) can be prepared by reacting a compound (25) with an alcohol, R³ OH, in an inert solvent in the presence of a condensing agent, and the reaction is conducted in a manner similar to that of process 2 in Step 3.

(Step 25)

In this step, a compound (27) can be prepared by reacting a compound (25) with an amine, R⁴ R⁵ NH, in an inert solvent in the presence of a condensing agent, and the reaction is conducted in a manner similar to that of Step 14.

(Step 26)

In this step, a compound (1B) can be prepared by removing the protecting groups, A and B⁶ in a compound (26) in an inert solvent, and the reaction is conducted in a manner similar to that of Step 22.

(Step 27)

In this step, a compound (1C) can be prepared by removing the protecting groups, A and B⁶ in a compound (27) in an inert solvent, and the reaction is conducted in a manner similar to that of Step 22.

(Step 28)

In this step, a compound (28) can be prepared by reacting a compound (25) with a compound (21), which is commercially available or prepared in Step 18, in an inert solvent, and the reaction is conducted in a manner similar to that of Step 25.

(Step 29)

In this step, a compound (29) can be prepared by reacting a compound (25) with a compound (19), which is commercially available or prepared in Step 16, in an inert solvent, and the reaction is conducted in a manner similar to that of Step 25.

(Step 30)

In this step, a compound (30) can be prepared by reacting a compound (25) with a compound (17), which is commercially available or prepared in Step 15, in an inert solvent, and the reaction is conducted in a manner similar to that of Step 25.

(Step 31)

In this step, a compound (1D) can be prepared by reacting a compound (28) with a reagent for deprotection in an inert solvent, and the reaction is conducted in a manner similar to that of Step 22.

(Step 32)

In this step, a compound (1E) can be prepared by reacting a compound (29) with a reagent for deprotection in an inert solvent, and the reaction is conducted in a manner similar to that of Step 22.

(Step 33)

In this step, a compound (1F) can be prepared by reacting a compound (30) with a reagent for deprotection in an inert solvent, and the reaction is conducted in a manner similar to that of Step 22.

EFFECT OF INVENTION

(Test Example 1)

Inhibitory activity on type IV collagenases

The activity of type IV collagenase was assayed according to the method of Salo et al. [J. Biol. Chem., Vol. 258, 3058-3063 (1983)].

In detail, the activity was assayed by measuring the cleavage of collagen by using type IV collagenases prepared from a serum-free culture medium of human melanoma cells, and type IV collagen prepared from mouse EHS tumor radio-labelled as a substrate.

The inhibitory activity on type IV collagenases was measured by calculating the inhibition percent of the enzyme reaction, in which the test samples were added to the reaction mixture.

The results (expressed as I₅₀) is shown below.

                  TABLE 2                                                          ______________________________________                                         Compound No.   IC.sub.50 (nmol/ml)                                             ______________________________________                                         Example 3      0.33                                                            Example 4      0.35                                                            Example 5      0.36                                                            Example 14     0.082                                                           Example 16     0.042                                                           Example 17     0.27                                                            Example 18     0.027                                                           Example 19     0.080                                                           Example 20     0.073                                                           Example 21     0.28                                                            Example 27     0.16                                                            Example 29     0.25                                                            Example 30     0.24                                                            Example 32     0.75                                                            Example 33     0.064                                                           Example 34     0.037                                                           Example 35     0.20                                                            Example 108    1.4                                                             Example 109    4.3                                                             Example 110    18.5                                                            ______________________________________                                    

POSSIBLE USEFULNESS IN INDUSTRY

The compounds of the present invention have excellent inhibitory activity on type IV collagenases, and are useful as inhibitors of angiogenesis, as inhibitors of cancer invasion or as inhibitors of cancer metastasis.

When the compounds of the present invention are employed as inhibitors of angiogenesis, as inhibitors of cancer invasion or as inhibitors of cancer metastasis, these compounds may be administered in various forms. As the mode of administration, there may be mentioned for example: oral administration by tablets, capsules, powders, granules or syrups; or parenteral administration by injections (intravenous, intramuscular, subcutaneous), eye drops or suppositories. These drug forms can be prepared according to conventional means, where the main ingredient is added with any known additives usually employable in the technological field of pharmaceutical preparation, such as vehicles, binders, disintegrators, lubricants, corrigents, solubilizers, suspending agents and/or coating agents. Though the dosage may be varied depending on patient's symptom, age, body weight, administration route and preparation forms, usually the amount from 50 mg to 1000 mg can be given to one adult a day.

BEST EMBODIMENT FOR WORKING THE INVENTION

The present invention is illustrated in detail by the following Examples, Referential Examples and Preparation Examples.

EXAMPLE 1 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic (4S)-5-methyl-3-oxohexan-4-ylamide

N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyamino-carbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (4S)-5-methyl-3-oxohexan-4-ylamide (75 mg), prepared in Referential Example 16, in methanol (3.0 ml) was catalytically reduced in the presence of 10% palladium on charcoal (8 mg) under an atmosphere of hydrogen at room temperature with stirring for 1.8 hr. After completion of the reaction, the catalyst was filtered off and the filtrate was concentrated under reduced pressure. The residue was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 10:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (28 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.77 (3H, d, J=6.6 Hz), 0.84 (3H, t, J=7.3 Hz), 0.93 (3H, d, J=6.6 Hz), 1.09 (3H, t, J=7.3 Hz), 1.13-2.68 (15H, complex), 2.57 (2H, q, J=7.3 Hz), 2.70-3.18 (2H, complex), 3.98 (1H, br.d, J=5.9 Hz), 4.64 (1H, dd, J=8.6, 4.6 Hz), 4.85 (1H, d, J=12.5 Hz), 5.37 (1H, br.s), 7.64 (1H, d, J=8.6 Hz), 9.91 (1H, m).

IR absorption spectrum (liquid film) cm⁻¹ : 3300 (m), 2940 (m), 1715 (m), 1660 (s), 1625 (s)

High resolution MS spectrum: [M]⁺ =426.2851 (C₂₁ H₃₈ N₄ O₅); Calcd. value: 426.2842 [α]_(D) ²⁶ =-30.60° (c=1.00, EtOH)

EXAMPLE 2 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (4S)-6-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (4S)-6-methyl-3-oxoheptan-4-ylamide (24 mg), prepared in Referential Example 23, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (10 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.78-0.98 (9H, complex), 1.09 (3H, t, J=6.9 Hz), 1.12-2.11 (15H, complex), 2.18-2.50 (2H, m), 2.59 (2H, m) , 2.70-3.12 (2H, complex), 4.00 (1H, m), 4.65 (1H, br.d, J=5.3 Hz), 4.87 (1H, br.d, J=11.9 Hz), 5.28 (1H, br.s), 7.68 (1H, br.d, J=5.3 Hz), 9.90 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3300 (m), 2945 (m), 1720 (m), 1660 (s), 1625 (s)

High resolution MS spectrum: [M+H]⁺ =441.3069 (C₂₂ H₄₁ N₄ O₅); Calcd. value: 441.3077 [α]_(D) ²⁶ =-32.5° (c=1.00, CHCl₃)

EXAMPLE 3 2-(S)-[N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazyl]aminoisovaleric acid N-methyl-N-methoxyamide

Following the procedure described in Example 1, the protecting groups of 2-(S)-[N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazyl]aminoisovaleric acid N-methyl-1-N-methoxyamide (39 mg), prepared in Referential Example 24, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 17:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and nethanol as an eluent, to give the desired compound (18 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.69-1.06 (9H, complex), 1.10-2.44 (14H, complex), 2.57 (1H, t, J=12.5 Hz), 2.82 (1H, m), 3.00 (1H, br.d, J=12.5 Hz), 3.26 (3H, s), 3.85 (3H, s), 3.97 (1H, br.d, J=6.6 Hz), 4.95 (1H, br.t, J=7.5 Hz), 5.08 (1H, d, J=11.9 Hz), 5.47 (1H, s), 8.16 (1H, br.d, J=8.6 Hz), 10.6 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3300 (m), 2945 (m), 1630 (s)

High resolution MS spectrum: [M]⁺ =457.2868 (C₂₁ H₃₉ N₅ O₆); Calcd. value: 457.2836 [α]_(D) ²⁶ =-43.8° (c=1.01, EtOH)

EXAMPLE 4 Methyl 2-(S)-[N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazyl]aminoisovalerate

Following the procedure described in Example 1, the protecting groups of methyl 2-(S)-[N¹ -benzyloxy-carbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazinyl]aminoisovalerate (49 mg), prepared in Referential Example 25, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×10 cm size, 0.5 mm thick), using a 20:1 mixture of chloroform and methanol as a developing solvent twice and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (20 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.67-1.02 (9H, complex), 1.04-2.60 (15H, complex), 2.82 (1H, m), 3.01 (1H, br.d, J=12.5 Hz), 3.77 (3H, s), 3.97 (1H, m), 4.57 (1H, dd, J=7.9 and 5.3 Hz), 4.92 (1H, d, J=11.9 Hz), 5.38 (1H, s), 7.74 (1H, br.d, J=7.3 Hz), 9.82 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3300 (m), 2940 (m), 1730 (m), 1660 (s), 1625 (s)

High resolution. MS spectrum: [M]⁺ =428.2631 (C₂₀ H₃₆ N₄ O₆); Calcd. value: 428.2635 [α]_(D) ²⁶ =-27.0° (c=1.02 EtOH)

EXAMPLE 5 tert-Butyl 2-(S)-[N² -[2-(R)-Hydroxyaminocarbonyl)-methyl-1-oxoheptyl]-(S)-piperazyl)aminoisovalerate

Following the procedure described in Example 1, the protecting groups of tert-butyl 2-(S)-[N¹ -benzyloxy-carbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazyl]aminoisovalerate (42 mg), prepared in Referential Example 26, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×10 cm size, 0.5 mm thick), using a 20:1 mixture of chloroform and methanol as a developing solvent twice and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (16 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.72-0.97 (9H, complex), 1.11-2.38 (14H, complex), 1.48 (9H, s), 2.49 (1H, dd, J=12.5 and 11.2 Hz), 2.83 (1H, m), 3.01 (1H, br.d, J=11.9 Hz), 3.96 (1H, m), 4.47 (1H, m), 4.89 (1H, d, J=11.2 Hz), 5.38 (1H, br.s), 7.54 (1H, br.d, J=7.9 Hz), 9.86 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3320 (m), 2945 (m), 1720 (w), 1665 (m), 1630 (s)

High resolution MS spectrum: [M]⁺ =470.3110 (C₂₃ H₄₂ N₄ O₆); Calcd. value: 470.3104 [α]_(D) ²⁶ =36.6° (c=1.00, EtOH)

EXAMPLE 6 N² -[2-(R)-hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid tert-butyl ester

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid tert-butyl ester (34 mg), prepared in Referential Example 29, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 20:1 mixture of chloroform and methanol as a developing solvent twice and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (14 mg) as colorless crystals having a m.p. of 110°-111° C. after recrystallization from a mixture of hexane and ethyl acetate.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.12-2.04 (11H, complex), 1.48 (9H, s), 2.18-2.49 (3H, complex), 2.71 (1H, m), 3.07 (1H, br.d, J=13.2 Hz), 4.00 (1H, m), 4.16 (1H, d, J=12.5 Hz), 5.19 (1H, d, J=4.0 Hz), 9.03 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3240 (m), 2940 (s), 1725 (s), 1630 (s)

High resolution MS spectrum: [M]⁺ =371.2397 (C₁₈ H₃₃ N₃ O₅); Calcd. value: 371.2419 [α]_(D) ²⁶ =+30.4° (c=0.50, EtOH)

EXAMPLE 7 N² -(3-hydroxyaminocarbonylpropionyl)-(S)-piperazic acid tert-butyl ester

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -(3-benzyloxyaminocarbonylpropionyl)-(S)-piperazic acid tert-butyl ester (84 mg), prepared in Referential Example 35, were removed by catalytic reduction. After working-up in a similar manner to that of Example 1, the desired compound (36 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.41 (1H, m, overlap to 6 1.48), 1.48 (9H, s), 1.61 (1H, m), 1.86 (1H, m), 2.18 (1H, br.d, J=13.2 Hz), 2.33-2.58 (2H, cpmplex), 2.66-3.12 (4H, complex), 4.28 (1H, br.d, J=12.5 Hz), 5.14 (1H, dd, J=4.6 and 1.3 Hz), 7.6-8.6 (1H, br.s), 9.59 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3250 (m), 2945 (m), 1725 (s), 1640 (s)

High resolution MS spectrum: [M]⁺ =301.1631 (C₁₃ H₂₃ N₃ O₅); Calcd. value: 301.1636 [α]_(D) ²⁶ =-12.9° (c=1.01 EtOH)

EXAMPLE 8 N² -(3-Hydroxyaminocarbonylpropionyl)-(S)-piperazic acid (4S,5S)-5-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -(3-benzyloxyaminocarbonylpropionyl)-(S)-piperazic acid (4S,5S)-5-methyl-3-oxoheptan-4-ylamide (111 mg), prepared in Referential Example 37, were removed by catalytic reduction. The product was purified by preparative reverse phase thin layer chromatography through silica gel (20×20 cm size, 0.25 mm thick, two plates), using a 1:1 mixture of water and methanol as an eluent, to give the desired compound (53 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=7.3 Hz), 0.92 (3H, d, J=6.6 Hz), 1.07 (3H, t, J=7.3 Hz), 1.28 (1H, m), 1.42-2.23 (6H, complex), 2.34-2.52 (2H, complex), 2.55 (2H, q, J=7.3 Hz), 2.75 (1H, m), 2.85-3.13 (3H, complex), 4.58 (1H, dd, J=7.9 and 5.3 Hz), 4.64 (1H, d, J=12.5 Hz), 5.27 (1H, d, J=4.6 Hz), 7.31 (1H, m), 8.12-8.83 (1H, br.s), 9.83 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3280 (s), 2965 (s), 1715 (s), 1635 (s)

High resolution MS spectrum: [M+H-OH]⁺ =354.2261 (C₁₇ H₃₀ N₄ O₄); Calcd. value: 354.2266 [α]_(D) ²⁶ =-12.2° (c=1.97, CHCl₃)

EXAMPLE 9 N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid methyl ester

To a solution of N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (34 mg), prepared in Referential Example 13, in ethyl acetate (2.0 ml) was added an ethereal solution of diazomethane with ice-cooling, until the evolution of nitrogen gas ceased. After completion of the reaction, the solvent was distilled off under reduced pressure. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 30:1 mixture of chloroform and methanol as a developing solvent to give a compound (11 mg). Following the procedure described in Example 1, the protecting groups of the compound thus obtained were removed by catalytic reduction and the resulting product was purified by preparative thin layer chromatography through silica gel (20×10 cm size, 0.5 mm thick), using a 25:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethylacetate and methanol as an eluent, to give the desired compound (4.2 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.83 (3H, t, J=6.6 Hz), 1.10-2.01 (11H, complex), 2.22 (1H, m), 2.30 (1H, dd, J=14.2 and 3.6 Hz), 2.54 (1H, dd, J=14.2 and 11.2 Hz), 2.73-3.14 (2H, complex), 3.77 (3H, s), 3.95 (1H, m), 4.20 (1H, d, J=11.9 Hz), 5.34 (1H, d, J=4.0 Hz), 7.35-8.20 (1H, m), 9.18 (1H, m)

IR absorption spectrum (film) cm⁻¹ : 3245 (m), 2950 (s), 1735 (s), 1630 (s)

High resolution MS spectrum: [M]⁺ =329.1965 (C₁₅ H₂₇ N₃ O₅); Calcd. value: 329.1950 [α]_(D) ²⁶ -15° (c=0.35, EtOH)

EXAMPLE 10 N² -[2-(R)-(oxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid tert-butyl ester (127 mg), prepared in Referential Example 12, were removed by catalytic reduction. After working up in a manner similar to that of Example 1, the desired compound (63 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=6.6 Hz), 1.11-1.70 (10H, complex), 1.48 (9H, s), 1.89 (1H, m), 2.03-2.38 (2H, complex), 2.52 (1H, m), 2.82-3.13 (2H, complex), 3.98 (1H, br.s), 4.28 (1H, br.d, J=10.6 Hz), 5.20 (1H, s)

IR absorption spectrum (liquid film) cm⁻¹ : 3225 (m), 2940 (s), 1725 (s), 1640 (br.s)

High resolution MS spectrum: [M+H]⁺ =372.2517 (C₁₈ H₃₄ N₃ O₅); Calcd. value: 372.2499 [α]_(D) ²⁶ =-10.7° (c=1.00, EtOH)

EXAMPLE 11 N² -[2-(R)-(oxyaminocarbonyl)methyl-4-methyl-1-oxopentyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-4-methyl-1-oxopentyl]-(S)-piperazic acid tert-butyl ester (20 mg), prepared in Referential Example 46, were removed by catalytic reduction. The product was worked up in a manner similar to that of Example 1 and the desired compound was obtained (11 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.89 (3H, d, J=6.6 Hz), 0.93 (3H, d, J=6.6 Hz), 1.10-1.70 (4H, complex), 1.48 (9H, s), 1.88 (1H, m), 2.19 (1H, br.d, J=11.6 Hz), 2.30 (1H, br.d, J=11.6 Hz), 2.50 (1H, m), 2.71-3.12 (3H, complex), 4.03 (1H, br.d, J=5.9 Hz), 4.25 (1H, d, J=11.9 Hz), 5.20 (1H, br.s), 9.39 (1H, s)

IR absorption spectrum (film) cm⁻¹ : 3240 (s), 2960 (s), 1725 (s), 1630 (s)

High resolution MS spectrum: [M]⁺ =357.2270 (C₁₇ H₃₁ N₃ O₅); Calcd. value: 357.2264 [α]_(D) ²⁶ =-12.3° (c=0.51, EtOH)

EXAMPLE 12 N¹ -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid t-butyl ester (32 mg), prepared in Referential Example 58, were removed by catalytic reduction. After working up in a manner similar to that of Example 1, the desired compound (15 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.12-1.70 (16H, complex), 1.49 (9H, s), 1.88 (1H, m), 2.18 (1H, br.d, J=11.7 Hz), 2.30 (1H, br.d, J=11.7 Hz), 2.53 (1H, m), 2.75-3.36 (2H, complex), 3.93 (1H, m), 4.27 (1H, d, J=11.9 Hz), 5.20 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3230 (m), 2940 (s), 1725 (s), 1635 (s)

High resolution MS spectrum: [M]⁺ =413.2903 (C₂₁ H₃₉ N₃ O₅); Calcd. value: 413.2890 [α]_(D) ²⁶ =-11.9° (c=1.00, EtOH)

EXAMPLE 13 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N-methylamide

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N-methylamide (14 mg), prepared in Referential Example 60, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×10 cm size, 0.5 mm thick), using a 10:1 mixture of chloroform and methanol as a developing solvent twice and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (9 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.05-2.15 (18H, complex), 2.30 (1H, m), 2.53 (1H, m), 2.80 (3H, d, J=4.6 Hz), 2.83 (1H, m), 3.03 (1H, br.d, J=11.9 Hz), 3.84 (1H, m), 4.64 (1H, d, J=9.9 Hz), 5.05 (1H, br.s), 6.55 (br.s), 9.2-9.8 (1H, br.s)

IR absorption spectrum (film) cm⁻ : 3260 (s), 2930 (s), 1650 (s), 1625 (s)

High resolution MS spectrum: [M]⁺ =352.2466 (C₁₈ H₃₂ N₃ O₄); Calcd. value: 352.2466 [α]_(D) ²⁶ =-3.4° (c=0.82, EtOH)

EXAMPLE 14 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid N-methylamide

Following the procedure described in Example 1, the protecting groups of an N-methylamide compound, which was prepared by condensing methylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid (45 mg), prepared in Referential Example 70, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (10 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.5 Hz), 0.99-1.95 (13H, complex), 2.05 (1H, br.d, J=10.6 Hz), 2.29 (1H, dd, J=13.9 and 3.3 Hz), 2.52 (1H, br.t, J=13.9 Hz), 2.79 (3H, d, J=4.6 Hz), 2.81 (1H, overlap to 2.79 ppm), 3.02 (1H, d, J=13.2 Hz), 3.88 (1H, m), 4.67 (1H, d, J=11.9 Hz), 5.06 (1H, s), 6.75 (1H, d, J=4.6 Hz), 9.52-9.91 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3271 (m), 2929 (s), 1648 (s), 1626 (s)

Mass spectrum [M-H₂ O]⁺ =324

High resolution MS spectrum: [M]⁺ =342.2256 (C₁₆ H₃₀ N₄ O₄); Calcd. value: 342.2267 [α]_(D) ²⁶ =-9.3° (c=0.90 EtOH)

EXAMPLE 15 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid N,N-dimethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-dimethylamide compound, which was prepared by condensing dimethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyamino-carbonyl)methyl-1-oxooctyl]-(S)-piperazic acid (41 mg), prepared in Referential Example 70, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (21 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=6.6 Hz), 1.12-2.07 (14H, complex), 2.29 (1H, dd, J=13.9 and 4.0 Hz), 2.53 (1H, dd, J=13.9 and 12.2 Hz), 2.70-3.19 (overlap to 2H, 3.06 ppm and 2.94 ppm), 2.94 (3H, s), 3.06 (3H, s), 3.92 (1H, m), 5.26 (1H, d, J=11.9 Hz), 5.51 (1H, br.s), 8.01-8.5 (1H, br.s), 9.41-9.70 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3249 (m), 2929 (s), 1645 (s), 1625 (s)

Mass spectrum [M]⁺ =356

High resolution MS spectrum: [M+H]⁺ =357.2511 (C₁₇ H₃₃ N₄ O₄); Calcd. value: 357.2502 [α]_(D) ²⁶ =+5.9° (c=1.0, EtOH)

EXAMPLE 16 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid N-methylamide

Following the procedure described in Example 1, the protecting groups of an N-methylamide compound, which was prepared by condensing methylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid (46 mg), prepared in Referential Example 80, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (20 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.00-1.95 (1H, complex), 2.05 (1H, br.d, J=3.3 Hz), 2.28 (1H, br.d, J=3.3 Hz), 2.51 (1H, dd, J=13.9 and 11.2 Hz), 2.78 (3H, d, J=4.6 Hz), 2.82 (1H, m), 3.02 (1H, br.d, J=12.5 Hz), 3.87 (1H, m), 4.73 (1H, d, J=11.9 Hz), 5.06 (1H, br.s), 6.93 (1H, m), 9.91-10.12 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3268 (m), 2928 (s), 1652 (s), 1626 (s)

Mass spectrum [M+H]⁺ =357

High resolution MS spectrum: [M+H]⁺ =357.2493 (C₁₇ H₃₃ N₄ O₄); Calcd. value: 357.2502 [α]_(D) ²⁶ =-9.5° (c=1.0, EtOH)

EXAMPLE 17 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid N,N-dimethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-dimethylamide compound, which was prepared by condensing dimethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid (49 mg), prepared in Referential Example 80, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (10 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.83 (3H, t, J=6.6 Hz), 1.05-1.78 (14H, complex), 1.80-2.04 (2H, complex), 2.26 (1H, br.d, J=12.5 Hz), 2.50 (1H, br. t, J=12.5 Hz), 2.72-3.12 (2H, complex), 2.93 (3H, s), 3.05 (3H, s), 3.90 (1H, m), 5.24 (1H, d, J=11.9 Hz), 5.53 (1H, m)

IR absorption spectrum (film) cm⁻¹ : 3260 (m), 2940 (s), 1625 (s)

Mass spectrum [M+H]⁺ =371

High resolution MS spectrum: [M+H]⁺ =371.2677 (C₁₈ H₃₅ N₄ O₄); Calcd. value: 371.2658 [α]_(D) ²⁶ =+7.20 (c=0.81, EtOH)

EXAMPLE 18 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid N-methylamide

Following the procedure described in Example 1, the protecting groups of an N-methylamide compound, which was prepared by condensing methylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid (52 mg), prepared in Referential Example 90, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (7 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.04-2.12 (22H, complex), 2.19-2.63 (2H, complex), 2.79 (3H, d, J=3.3 Hz), 2.80 (1H, m), 3.03 (1H, br.d, J=13.2 Hz), 3.85 (1H, m), 4.64 (1H, br.d, J=11.9 Hz), 5.05 (1H, br.s), 6.68 (1H, m), 9.48-9.76 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3267 (m), 2855 (s), 1652 (s), 1625 (s)

Mass spectrum [M]⁺ =398

High resolution MS spectrum: [M-H₂ O]⁺ =380.2809 (C₂₀ H₃₆ N₄ O₃); Calcd. value: 380.2787 [α]_(D) ²⁶ =-8.9° (c=0.61, EtOH)

EXAMPLE 19 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid N,N-dimethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-dimethylamide compound, which was prepared by condensing dimethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid (50 mg), prepared in Referential Example 90, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (27 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.06-1.78 (20H, complex), 1.80-2.02 (2H, complex), 2.29 (1H, dd, J=14.5 and 4.0 Hz), 2.52 (1H, dd, J=14.5 and 10.9 Hz), 2.70-3.12 (2H, complex), 2.94 (3H, s), 3.06 (3H, s), 3.92 (1H, m), 5.26 (1H, d, J=11.9 Hz), 5.50 (1H, br.s), 7.92-8.77 (1H, br.m), 9.59 (1H, m)

IR absorption spectrum (film) cm⁻¹ : 3262 (m), 2926 (s), 1649 (s), 1627 (s)

Mass spectrum [M]⁺ =412

High resolution MS spectrum: [M]⁺ 412.3036 (C₂₁ H₄₀ N₄ O₄); Calcd. value: 412.3049 [α]_(D) ²⁶ =+6.8° (c=1.0, EtOH)

EXAMPLE 20 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid N-methylamide

Following the procedure described in Example 1, the protecting groups of an N-methylamide compound, which was prepared by condensing methylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid (45 mg), prepared in Referential Example 100, were removed by catalytic reduction. The product was purified by preparative thin layer chromatograpky through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (17 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.37-2.09 (6H, complex), 2.32 (1H, dd, J=14.2 and 4.3 Hz), 2.43-2.90 (4H, complex), 2.76 (3H, d, J=4.6 Hz), 2.97 (1H, d, J=13.9 Hz), 3.92 (1H, m), 4.74 (1H, d, J=12.0 Hz), 5.07 (1H, d, J=2.1 Hz), 6.82 (1H, br.d, J=4.0 Hz), 7.05-7.31 (5H, complex), 9.69-10.10 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3440 (m), 3270 (m), 2939 (m), 1640 (s), 1629 (s)

Mass spectrum [M+H]⁺ =363

High resolution MS spectrum: [M]⁺ =362.1947 (C₁₈ H₂₆ N₄ O₄); Calcd. value: 362.1954 [α]_(D) ²⁶ =+1.0° (c=1.0, EtOH)

EXAMPLE 21 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid N,N-dimethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-dimethylamide compound, which was prepared by condensing dimethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid (48 mg), prepared in Referential Example 100, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (9 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.37-2.04 (6H, complex), 2.32 (1H, m), 2.48-2.69 (3H, complex), 2.71-3.12 (2H, complex), 2.92 (3H, s), 3.03 (3H, s), 3.97 (1H, m), 5.26 (1H, br.d, J=12.5 hz), 5.52 (1H, br.d, J=2.6 Hz), 7.08-7.30 (5H, complex)

IR absorption spectrum (film) cm⁻¹ : 3250 (m), 2925 (m), 1625 (s)

Mass spectrum [M+Me]⁺ =361

High resolution MS spectrum: [M]⁺ =376.2126 (C₁₉ H₂₈ N₄ O₄); Calcd. value: 376.2110 [α]_(D) ²⁶ =+19.8° (c=0.71, EtOH)

EXAMPLE 22 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N,N-dimethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-dimethylamide compound, which was prepared by condensing dimethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (50 mg), prepared in Referential Example 13, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (25 mg).

NMR spectrum (270 MHz, CDCl₃) δppm: 0.85 (3H, t, J=6.6 Hz), 1.12-2.09 (12H, complex), 2.38 (1H, dd, J=14.2 and 4.0 Hz), 2.53 (1H, dd, J=14.2 and 10.8 Hz), 2.70-3.13 (2H, complex), 2.93 (3H, s), 3.05 (3H, s), 3.92 (1H, m), 5.26 (1H, d, J=11.9 Hz), 5.51 (1H, br.s), 9.28-9.92 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3265 (m), 2990 (s), 1635 (s), 1625 (s)

Mass spectrum [M]⁺ =342

High resolution MS spectrum: [M]⁺ =342.2284 (C₁₆ H₃₀ N₄ O₄); Calcd. value: 342.2267

EXAMPLE 23 N² -(4-Hydroxyamino-1,4-dioxobutyl)-(S)-piperazic acid N,N-dimethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-dimethylamide compound, which was prepared by condensing dimethylamine with N¹ -benzyloxycarbonyl-N² -(3-benzyloxyaminocarbonylpropionyl)-(S)-piperazic acid (70 mg), prepared in Referential Example 36, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (25 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.47-1.79 (2H, complex), 1.82-2.03 (2H, complex), 2.29-2.56 (2H, complex), 2.62-3.18 (4H, complex), 2.94 (3H, s), 3.07 (3H, s), 5.18 (1H, d, J=11.9 Hz), 5.45 (1H, t, J=4.0 Hz), 9.50-9.89 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3260 (m), 2942 (m), 1630 (s)

Mass spectrum [M]⁺ =272

High resolution MS spectrum: [M]⁺ =272.1471 (C₁₁ H₂₀ O₄ N₄); Calcd. value: 272.1484 [α]_(D) ²⁶ =+4.0° (c=1.0 EtOH)

EXAMPLE 24 N² -(4-Hydroxyamino-1,4-dioxobutyl)-(S)-piperazic acid N-methylamide

Following the procedure described in Example 1, the protecting groups of an N-methylamide compound, which was prepared by condensing methylamine with N¹ -benzyloxycarbonyl-N² -(3-benzyloxyaminocarbonylpropionyl)-(S)-piperazinecarboxylic acid (82 mg), prepared in Referential Example 36, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (24 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.41-1.65 (2H, complex), 1.80 (1H, m), 2.21 (1H, br.d, J=13.1 Hz), 2.37 (2H, t, J=7.3 Hz), 2.65-2.88 (2H, complex), 2.76 (3H, s), 2.90-3.01 (2H, complex), 5.03 (1H, dd, J=4.0 and 3.0 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 3265 (m), 2941 (m), 1641 (s)

Mass spectrum [M]⁺ =258

High resolution MS spectrum: [M]⁺ =258.1332 (C₁₀ H₁₈ N₄ O₄); Calcd. value: 258.1328 [α]_(D) ²⁶ =-33.5° (c=1.0, EtOH)

EXAMPLE 25 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid pyrrolidinylamide

Following the procedure described in Example 1, the protecting groups of a pyrrolidinylamide compound, which was prepared by condensing pyrrolidine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (42 mg), prepared in Referential Example 13, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (15 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=6.6 Hz), 1.01-2.08 (16H, m), 2.30 (1H, dd, J=13.9 and 4.0 Hz), 2.53 (1H, dd, J=13.9 and 10.9 Hz), 2.83 (1H, m), 3.03 (1H, br.d, J=13.2 Hz), 3.29-3.71 (4H, m), 3.91 (1H, m), 5.30 (1H, m), 5.32 (1H, br.s), 7.88-8.44 (1H, br.s), 9.28-9.70 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3250 (m), 2940 (s), 1655 (m), 1620 (s), 1625 (s)

High resolution MS spectrum: [M]⁺ =368.2404 (C₁₈ H₃₂ N₄ O₄); Calcd. value: 368.2424

EXAMPLE 26 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N,N-diethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-diethylamide compound, which was prepared by condensing diethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (42 mg), prepared in Referential Example 13, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (19 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.85 (3H, t, J=6.6 Hz), 1.11 (3H, t, J=6.9 Hz), 1.13-2.08 (15H, complex), 2.29 (1H, dd, J=14.5 and 4.0 Hz), 2.52 (1H, dd, J=14.5 and 11.0 Hz), 2.81 (1H, dd, J=13.9 and 11.9 Hz), 3.04 (1H, br.d, J=13.9 Hz), 3.08-3.47 (3H, complex), 3.53 (1H, m), 3.90 (1H, m), 5.32 (1H, d, J=11.2 Hz), 5.43 (1H, d, J=5.3 Hz), 7.97-8.62 (1H, br.s), 9.25-9.69 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3255 (m), 2940 (m), 1620 (s)

Mass spectrum [M]⁺ =370

High resolution MS spectrum: [M]⁺ =370.2584 (C₁₈ H₃₄ N₄ O₄); Calcd. value: 370.2580

EXAMPLE 27 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N-ethylamide

Following the procedure described in Example 1, the protecting groups of an N-ethylamide compound, which was prepared by condensing ethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (45 mg), prepared in Referential Example 13, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (19 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=6.6 Hz), 1.12 (3H, t, J=7.3 Hz), 1.15-1.95 (11H, complex), 2.05 (1H, m), 2.28 (1H, dd, J=13.9 and 3.3 Hz), 2.51 (1H, dd, J=13.9 and 11.2 Hz), 2.85 (1H, m), 3.02 (1H, br.d, J=12.5 Hz), 3.26 (2H, m), 3.91 (1H, m), 4.72 (1H, d, J=12.5 Hz), 5.05 (1H, s), 6.74 (1H, br.s), 9.73-10.12 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3255 (m), 2910 (m), 1645 (s), 1605 (s)

Mass spectrum [M-NHEt]⁺ =298

High resolution MS spectrum: [M-NHEt]⁺ =298.1771 (C₁₄ H₂₄ N₃ O₄); Calcd. value: 298.1767

EXAMPLE 28 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid piperazylamide

Following the procedure described in Example 1, the protecting groups of an N-benzylpiperazinylamide compound, which was prepared by condensing N-benzylpiperazine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (43 mg), prepared in Referential Example 13, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a. 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol,as an eluent, to give the desired compound (6 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=6.6 Hz), 1.12-1.77 (10H, complex), 1.79-2.02 (2H, complex), 2.20-2.63 (7H, complex), 2.80 (1H, m), 3.02 (1H, br.d, J=13.2 Hz), 3.32-3.78 (4H, complex), 3.88 (1H, m), 5.17 (1H, d, J=11.5 Hz), 5.50 (1H, dd, J=5.6 and 2.3 Hz), 9.00-9.44 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3250 (m), 2920 (s), 1630 (s), 1620 (s)

Mass spectrum [M-H₂ O]⁺ =365

EXAMPLE 29 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazylamide

Following the procedure described in Example 1, the protecting groups of an amide compound, which was prepared by condensing ammonia and N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl)-(S)-piperazic acid (46 mg), prepared in Referential Example 13 with ammonia, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (10 mg).

NMR spectrum (270 MHz, CD₃ OD) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.17-1.78 (10H, complex), 1.97 (1H, m), 2.11 (1H, m), 2.15 (1H, dd, J=13.9 and 5.9 Hz), 2.38 (1H, dd, J=13.9 and 9.2 Hz), 2.78-3.08 (2H, complex), 3.94 (1H, m), 5.13 (1H, br.d, J=5.1 Hz)

IR absorption spectrum (KBr, pellet): 3267 (m), 2930 (s), 1683 (s), 1628 (s)

Mass spectrum [M-NH₂ ]⁺ =298

High resolution MS spectrum: [M-NH₂ ]⁺ =298.1758 (C₁₄ H₂₄ N₃ O₄); Calcd. value: 298.1766 [α]_(D) ²⁶ =+6.4° (c=0.41, EtOH)

EXAMPLE 30 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N-isobutylamide

Following the procedure described in Example 1, the protecting groups of an N-isobutylamide compound, which was prepared by condensing isobutylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (42 mg), prepared in Referential Example 13, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm Size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (18 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=6.6 Hz), 0.89 (6H, d, J=6.6 Hz), 1.04-2.09 (13H, complex), 2.23 (1H, m), 2.47 (1H, m), 2.69-3.08 (3H, complex), 3.16 (1H, m), 3.88 (1H, m), 4.75 (1H, br.d, J=10.6 Hz), 5.09 (1H, br.s), 7.01 (1H, m)

IR absorption spectrum (KBr, pellet): 3276 (m), 2958 (s), 2931 (s), 1645 (s), 1629 (s)

Mass spectrum [M-H₂ O]⁺ =352

High resolution MS spectrum: [M]⁺ =370.2583 (C₁₈ H₃₄ O₄ N₄); Calcd. value: 370.2580

EXAMPLE 31 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid t-butyl ester

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid tert-butyl ester (32 mg), prepared in Referential Example 69, were removed by catalytic reduction. The product was,purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (17 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.16-1.70 (12H, complex), 1.48 (9H, s), 1.86 (1H, m), 2.19 (1H, br.d, J=14.4 Hz), 2.32 (1H, dd, J=13.9 and 3.7 Hz), 2.56 (1H, br.dd, J=13.9 and 11.0 Hz), 2.84 (1H, br.t, J=15.0 Hz), 3.01 (1H, br.d, J=14.4 Hz), 3.90 (1H, m), 4.15-4.47 (1H, br.s), 5.20 (1H, d, J=3.9 Hz).

IR absorption spectrum (film) cm⁻¹ : 3233 (m), 2931 (s), 1728 (s), 1633 (s)

Mass spectrum [M]⁺ =385

High resolution MS spectrum: [M]⁺ =385.2576 (C₁₉ H₃₅ N₃ O₅); Calcd. value: 385.2577 [α]_(D) ²⁶ =-11.4° (c=1.0, EtOH)

m.p. 61°-63° C.

EXAMPLE 32 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxo-4-methyl-pentyl]-(S)-piperazic acid N,N-dimethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-dimethylamide compound, which was prepared by condensing dimethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxo-4-methylpentyl]-(S)-piperazic acid (49 mg), prepared in Referential Example 101, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (20 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 0.92 (3H, d, J=6.6 Hz), 1.20 (1H, m), 1.38-1.78 (4H, complex), 1.80-2.04 (2H, complex), 2.28 (1H, dd, J=13.9 and 4.0 Hz), 2.49 (1H, dd, J=13.9 and 11.2 Hz), 2.70-3.17 (2H, complex, overlap to 3.06 ppm and 2.94 ppm), 2.94 (3H, s), 3.06 (3H, s), 4.04 (1H, m), 5.26 (1H, d, J=11.2 Hz), 5.52 (1H, br.s), 8.15-8.61 (1H, br.s), 9.42-9.74 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3363 (m), 2954 (s), 1625 (s)

Mass spectrum [M+H]⁺ =329

High resolution MS spectrum: [M+H]⁺ =329.2195 (C₁₅ H₂₉ N₄ O₄); Calcd. value: 329.2189 [α]_(D) ²⁶ =+3.1° (c=1.0, EtOH)

EXAMPLE 33 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N,N-dimethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-dimethylamide compound, which was prepared by condensing dimethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid (49 mg), prepared in Referential Example 59, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (25 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.02-1.78 (16H, complex), 1.81-2.04 (2H, complex), 2.29 (1H, dd, J=14.5 and 4.0 Hz), 2.52 (1H, dd, J=14.5 and 10.9 Hz), 2.70-3.11 (2H, complex), 2.94 (3H, s), 3.06 (3H, s), 3.93 (1H, m), 5.26 (1H, d, J=11.2 Hz), 5.51 (1H, br.s), 8.09-8.62 (1H, br.s), 9.50-9.72 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3250 (w), 2910 (s), 1635 (s), 1625 (s)

Mass spectrum [M]⁺ =384

High resolution MS spectrum: [M]⁺ =384.2738 (C₁₉ H₃₆ N₄ O₄); Calcd. value: 384.2737

EXAMPLE 34 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N-ethylamide

Following the procedure described in Example 1, the protecting groups of an N-ethylamide compound, which was prepared by condensing ethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid (46 mg), prepared in Referential Example 59, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (20 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.12 (3H, t, J=7.3 Hz), 1.13-1.64 (15H, complex), 1.66-1.93 (2H, complex), .2.04 (1H, m), 2.28 (1H, br.dd, J=13.4 and 2.6 Hz), 2.51 (1H, br.t, J=12.5 Hz), 2.85 (1H, m), 3.02 (1H, br.d, J=12.5 Hz), 3.14-3.40 (2H, complex), 3.96 (1H, m), 4.71 (1H, d, J=11.9 Hz), 5.04 (1H, br.s), 6.73 (1H, br.s), 9.68-10.11 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3273 (m), 2928 (s), 1650 (s), 1626 (s)

Mass spectrum [M+H₂ O]⁺ =366

High resolution MS spectrum: [M+H₂ O]⁺ =366.2625 (C₁₉ H₃₄ N₄ O₃); Calcd. value: 366.2631 [α]_(D) ²⁶ =-5.4° (c=1.0, EtOH)

EXAMPLE 35 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N,N-diethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-diethylamide compound, which was prepared by condensing diethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid (47 mg), prepared in Referential Example 59, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (8 mg).

NMR spectrum (270 MHz; CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.12 (3H, t, J=6.6 Hz), 1.12-2.05 (21H, complex), 2.31 (1H, dd, J=13.9 and 3.6 Hz), 2.54 (1H, dd, J=13.9 and 10.6 Hz), 2.79 (1H, m), 3.22-3.44 (2H, complex), 3.04 (1H, br.d, J=13.9 Hz), 3.16 (1H, dq, J=13.9 and 6.9 Hz), 3.55 (1H, dq, J=14.5 and 7.3 Hz), 3.87 (1H, m), 5.32 (1H, d, J=11.9 Hz), 5.43 (1H, d, J=4.6 Hz), 7.40-8.03 (1H, br.s), 9.10-9.43 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3254 (s), 2856 (s), 1622 (s)

Mass spectrum [M]⁺ =412

High resolution MS spectrum: [M]⁺ =412.3034 (C₂₁ H₄₀ N₄ O₄); Calcd. value: 412.3050 [α]_(D) ²⁶ =-11.8° (c=0.39, EtOH)

EXAMPLE 36 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxo-4-methyl-pentyl]-(S)-piperazic acid N-methylamide

Following the procedure described in Example 1, the protecting groups of an N-methylamide compound, which was prepared by condensing methylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxo-4-methylpentyl]-(S)-piperazic acid (33 mg), prepared in Referential Example 101, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (11 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.89 (3H, d, J=5.9 Hz), 0.93 (3H, d, J=5.9 Hz), 1.22 (1H, m), 1.36-1.97 (5H, complex), 2.05 (1H, m), 2.28 (1H, dd, J=13.2 and 2.6 Hz), 2.49 (1H, dd, J=13.2 and 7.9 Hz), 2.77-3.11 (2H, complex), 2.78 (3H, d, J=4.0 Hz), 3.96 (1H, br.d, J=5.3 Hz), 4.71 (1H, d, J=6.9 Hz), 5.06 (1H, br.d, J=1.3 Hz), 6.85 (1H, br.s), 9.58-10.20 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3270, (m), 2960 (m), 1650 (s), 1625 (s)

Mass spectrum [M+H]⁺ =315

High resolution MS spectrum: [M+H]⁺ =315.2019 (C₁₄ H₂₇ N₄ O₄); Calcd. value: 315.2032 [α]_(D) ²⁶ =-10.6° (c=1.0, EtOH)

EXAMPLE 37 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid N-methylamide

Following the procedure described in Example 1, the protecting groups of an N-methylamide compound, which was prepared by condensing methylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid (59 mg), prepared in Referential Example 30, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (19 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.01-1.80 (11H, complex), 2.28 (1H, br.d, J=12.5 Hz), 2.33-2.90 (3H, complex), 2.77 (3H, br.s), 3.05 (1H, br.d, J=12.5 Hz), 3.99 (1H, m), 4.20 (1H, br.d, J=11.2 Hz), 5.11 (1H, br.s), 7.64 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3255 (m), 2929 (s), 1637 (s)

Mass spectrum [M]⁺ =328

High resolution MS spectrum: [M]⁺ =328.2108 (C₁₅ H₂₈ N₄ O₄); Calcd. value: 328.2110 [α]_(D) ²⁶ =+70.90 (c=1.0, EtOH)

EXAMPLE 38 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid N,N-dimethylamide

Following the procedure described in Example 1, the protecting groups of an N,N-dimethylamide compound, which was prepared by condensing dimethylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid (42 mg), prepared in Referential Example 30, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (18 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.14-1.75 (10H, complex), 1.81-2.05 (2H, complex), 2.28 (2H, d, J=7.3 Hz), 2.70 (1H, m), 2.95 (3H, s), 3.09 (3H, s), 3.11 (1H, m, overlap to 3.09 ppm), 4.10 (1H, m), 4.95 (1H, dd, J=11.2 and 1.3 Hz), 5.43 (1H, dd, J=5.3 and 3.3 Hz), 7.53-7.82 (1H, br.s), 8.68-8.92 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3261 (s), 2929 (s), 1622 (s)

Mass spectrum [M]⁺ =342

High resolution MS spectrum: [M]⁺ =342.2250 (C₁₆ H₃₀ N⁴ O₄); Calcd. value: 342.2267 [α]_(D) ²⁶ =+25.6° (c=1.0., EtOH)

EXAMPLE 39 N² -[2-(S)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N-methylamide

Following the procedure described in Example 1, the protecting groups of an N-methylamide compound, which was prepared by condensing methylamine with N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (41 mg), prepared in Referential Example 106, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (8 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.03-2.05 (11H, complex), 2.18-2.91 (4H, complex), 2.81 (3H, d, J=4.0 Hz), 3.06 (1H, d, J=13.2 Hz), 3.79-4.12 (2H, complex), 5.12 (1H, d, J=4.0 Hz), 7.37 (1H, d, J=4.0 Hz), 9.02-9.53 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3250 (m), 2930 (s), 1635 (s)

Mass spectrum [M-H₂ O]⁺ =310

High resolution MS spectrum: [M-H₂ O]⁺ =310.2006 (C₁₅ H₂₆ N₄ O₃); Calcd. value: 310.2005 [α]_(D) ²⁶ =-70.2° (c=0.28, EtOH)

EXAMPLE 40 N² -[2-(S)-(hydroxyaminocarbonyl)methyl-1-oxopentyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(S)-(benzyloxyaminocarbonyl)methyl-1-oxopentyl]-(S)-piperazic acid tert-butyl ester (40 mg), prepared in Referential Example 105, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 13:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (16 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.17-1.98 (11H, complex), 1.48 (9H, s), 2.05-2.50 (3H, complex), 2.72 (1H, dq, J=2.3 and 12.9 Hz), 3.08 (1H, dd, J=13.7 and 1.9 Hz), 4.00 (1H, m), 4.16 (1H, d, J=12.5 Hz), 5.19 (1H, d, J=4.0 Hz), 8.78-9.33 (1H, br.s)

IR absorption spectrum (film) cm⁻¹ : 3245 (m), 2940 (s), 1725 (s), 1650 (s), 1630 (s)

Mass spectrum [M]⁺ =371

High resolution MS spectrum: [M]⁺ =371.2408 (C₁₈ H₃₃ N₃ O₅); Calcd. value: 371.2421 [α]_(D) ²⁶ =-30.2° (c=0.48, EtOH)

Referential Example 1 N-Benzyloxycarbonyl-L-isoleucine (N-methyl-N-methoxy)-amide

A solution of N-benzyloxycarbonyl-L-isoleucine (15 g) in dichloromethane (200 ml) was cooled to 0° C., whereupon N,O-dimethylhydroxylamine hydrochloride (5.8 g), dicyclohexylcarbodiimide (DCC, 11.7 g), diisopropylethylamine (10 ml) and 4-dimethylaminopyridine (70 ml) were added successively thereto, and the resulting mixture was stirred at 0° C. for 2.3 hours. After filtering off precipitates, the reaction mixture was poured into hydrochloric acid and extracted with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over sodium sulfate. After the solvent had been distilled off under reduced pressure, the residue was purified by column chromatography through silica gel, using a 5:2 mixture of hexane and ethyl acetate as an eluent, to give 16.9 g of the desired compound as colorless crystals. Recrystallization from aqueous methanol gave rise to colorless crystals having a m.p. of 64°-66° C.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=7.3 Hz), 0.93 (3H, d, J=6.8 Hz), 1.12 (1H, m), .1.57 (1H, m), 1.73 (1H, m), 3.22 (3H, s), 3.79 (3H, s), 4.67 (1H, br.t, J=8.1 Hz), 5.06 (1H, d, J=12.5 Hz), 5.13 (1H, d, J=12.5 Hz), 5.35 (1H, br.d, J=9.8 Hz), 7.23-7.41 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3306 (m), 1719 (s), 1654 (s)

High resolution MS spectrum: [M+H]⁺ =309.1804 (C₁₆ H₂₅ N₂ O₄); Calcd. value: 309.1813 [α]_(D) ²⁶ =-4.68° (c=2.01, CHCl₃)

Referential Example 2 (4S,5S)-4-Benzyloxycarbonylamino-5-methyl-3-oxoheptane

A 0.99M tetrahydrofuran solution (16 ml) of ethylmagnesium bromide was added, dropwise, to a solution of N-benzyloxycarbonyl-L-isoleucyl (N-methyl-N-methoxy)amine (1.71 g) in tetrahydrofuran (40 ml) which had been Cooled to -15° C., under an atmosphere of nitrogen, and the resulting mixture was stirred for 0.6 hour and at room temperature for 0.6 hour. The reaction mixture was poured into a 5% aqueous solution of potassium hydrogensulfate and extracted with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over sodium sulfate. After the solvent had been distilled off under reduced pressure, the residue was purified by column chromatography through silica gel, using 4:1 and 2:1 mixtures of hexane and ethyl acetate as the eluent, to give 861 mg (56.2%) of the desired compound. From the eluate, N-benzyloxycarbonyl-L-isoleucyl (N-methyl-N-methoxy)-amine (508 mg), used as a starting material, was recovered. The desired compound was recrystallized from aqueous methanol to give colorless crystals having a m.p. of 57°-58° C.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=7.3 Hz), 0.98 (3H, d, J=6.8 Hz), 1.08 (3H, t, J=7.3 Hz), 1.27 (1H, m), 1.90 (1H, m), 2.52 (1H, dd, J=7.3 and 3.9 Hz), 4.36 (1H, dd, J=8.5 and 4.6 Hz), 5.09 (2H, s), 5.36 (1H, br.d, J=8.3 Hz), 7.24-7.40 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3270 (w), 1710 (s)

High resolution MS spectrum [M+H]⁺ =278.1750 (C₁₆ H₂₄ NO₃); Calcd. value: 278.1756

Referential Example 3 4-(S)-Isopropyl-3-(1-oxoheptyl)-2-oxazolidinone

A solution of 4-(S)-isopropyl-2-oxazolidinone (5.04 g) in tetrahydrofuran (125 ml) was cooled to -78° C. under an atmosphere Of nitrogen. n-Butyl lithium (16 ml, 1.65M, in hexane).was added dropwise to the solution. Heptanoyl chloride (6.4 ml) was added to the resulting mixture after 10 min stirring, and the stirring was continued for another 1.5 hours at the same temperature. The reaction mixture was then poured into a 5% aqueous solution of ammonium chloride and was extracted with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over sodium sulfate. After the solvent was distilled off under reduced pressure, the residue was purified by column chromatography through silica gel, using a 8:1 mixture of hexane and ethyl acetate as an eluent, to give the desired compound (9.69 g) as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.82-0.97 (3H, t, overlapped to 0.92 ppm), 0.87 (3H, d, J=6.8 Hz), 0.92 (3H, d, J=6.8 Hz), 1.22-1.45 (6H, complex), 1.58-1.75 (2H, complex), 2.38 (1H, d, hep, J=3.4 and 6.8 Hz), 2.77-3.06 (2H, complex), 4.19 (1H, dd, J=8.3 and 3.4 Hz), 4.26 (1H, t, J=8.3 Hz), 4.43 (1H, dt, J=8.3 and 3.4 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 1784 (s), 1703 (s)

High resolution MS spectrum [M]⁺ =241.1675 (C₁₃ H₂₃ NO₃); Calcd. value: 241.1677

Referential Example 4 4-(S)-Isopropyl-3-[2-(R)-tert-butoxycarbonylmethyl-1-oxoheptyl)-2-oxazolidinone

A solution of 4-(S)-isopropyl-3-(1-oxoheptyl)-2-oxazolidinone (519 mg) in tetrahydrofuran (15 ml) was cooled to -15° C., a 0.58M solution (39 ml) of lithium diisopropylamide in tetrahydrofuran was added thereto under an atmosphere of nitrogen, and the resulting mixture was stirred at -78° C for 10 minutes. A solution of tert-butyl bromoacetate (1.7 ml) in tetrahydrofuran (5 ml) was then added thereto and the temperature was allowed to rise gradually to -55° C. over a period of 5.5 hours with stirring. The reaction mixture was poured into a 5% aqueous solution of ammonium chloride and extracted with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over sodium sulfate. After the solvent was distilled off under reduced pressure, the residue was purified by column chromatography through silica gel using a 10:1 mixture of hexane and ethyl acetate as an eluent, to give 697 mg (91.2%) of the desired compound. Recrystallization from aqueous methanol gave rise to crystals having a m.p. of 51°-53° C.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=6.4 Hz), 0.91 (3H, d, J=6.3 Hz), 0.93 (3H, d, J=6.3 Hz), 1.14-1.51 (7H, complex), 1.41 (9H, s), 1.62 (1H, m), 2.38 (1H, d, hep, J=3.4 and 6.3 Hz), 2.43 (1H, dd, J=16.6 and 4.9 Hz), 2.74 (1H, dd, J=16.6 and 10.3 Hz), 4.15 (1H, m), 4.20 (1H, dd, J=7.9 and 3.4 Hz), 4.25 (1H, t, J=7.9 Hz), 4.43 (1H, dt, J=7.9 and 3.4 Hz)

IR absorption spectrum (KBr pellet) cm⁻¹ : 1763 (s), 1730 (s), 1702 (s)

High resolution MS spectrum [M+H]⁺ =356.2449 (C₁₉ H₃₄ NO₅); Calcd. value: 356.2437 [α]_(D) ²⁶ =+50.8° (c=1.03, CHCl₃)

Referential Example 5 2-(R)-(tert-Butoxycarbonylmethyl)heptanoic acid

A solution of 4-(S)-3-[2-(R)-(tert-butoxycarbonylmethyl)-1-oxoheptyl]-2-oxazolidinone (691 mg), prepared in Referential Example 4, in 40 ml of a 3:1 mixture of tetrahydrofuran and water was cooled to 0° C., and lithium hydroxide hydrate (165 mg) and a 31% aqueous solution of hydrogen peroxide (1.0 ml) were added successively thereto. The resulting mixture was stirred at 0° C. for 1.5 hours, then a 1.5N aqueous solution of sodium sulfite was added thereto. After several minutes stirring, the reaction mixture was poured into a 1N aqueous solution of sodium hydroxide followed by washing with dichloromethane. The pH of the aqueous layer was adjusted to pH 1 to 2 with 1N hydrochloric acid and the mixture was extracted with ethyl acetate. The extract was dried over sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography through silica gel, using a 8:1 mixture of hexane and ethyl acetate as an eluent, to give 452 mg (95.1%) of the desired compound as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=7.4 Hz), 1.21-1.41 (6H, complex), 1.43 (9H, s), 1.52 (1H, m), 1.65 (1H, m), 2.38 (1H, dd, J=16.5 and 5.3 Hz), 2.62 (1H, dd, J=16.5 and 9.2 Hz), 2.80 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 1734 (s), 1709 (s)

High resolution MS spectrum [M+H]⁺ =245.1752 (C₁₃ H₂₅ O₄); Calcd. value: 245.1752 [α]_(D) ²⁶ =+14.5° (c=1.97, EtOH)

Referential Example 6 tert-Butyl 3-(R)-benzyloxycarbonyloctanoate

Sodium hydrogencarbonate (305 mg) and benzyl bromide (1.0 ml) were added Successively to a solution of 2-(R)-(tert-butoxycarbonylmethyl)heptanoic acid (440 mg), prepared in Referential Example 5, in 18 ml of dimethylformamide (DMF), and the resulting mixture was stirred overnight at room temperature. The reaction mixture was poured into water and extracted with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over anhydrous sodium sulfate. After the solvent had been distilled off under reduced pressure, the residue was purified by column chromatography through silica gel, using a 20:1 mixture of hexane and ethyl acetate as an eluent, to give 476 mg of the desired compound as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.85 (3H, t, J=6.6 Hz), 1.17-1.32 (6H, complex), 1.41 (9H, s), 1.50 (1H, m), 1.61 (1H, m), 2.36 (1H, dd, J=16.5 and 5.3 Hz), 2.65 (1H, dd, J=16.5 and 9.2 Hz), 2.83 (1H, m), 5.09 (1H, d, J=12.5 Hz), 5.18 (1H, d, J=12.5 Hz), 7.24-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 1731 (s)

High resolution MS spectrum [M+H]⁺ =335.2230 (C₂₂ H₃₁ O₄); Calcd. value: 335.2223 [α]_(D) ²⁶ =+0.22° (c=7.9, CHCl₃)

Referential Example 7 3-(R)-Benzyloxycarbonyloctanoic acid

A 4M hydrochloric acid-dioxane solution (15 ml) was added to tert-butyl 3-(R)-benzyloxycarbonyloctanoate (983 mg), prepared in Referential Example 6, and the resulting mixture was stirred overnight. The reaction mixture was poured into water and extracted with ethyl acetate. The organic extract was dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography through silica gel, using a 30:1 mixture of chloroform and methanol as an eluent, to give 838 mg of the desired compound as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.85, (3H, t, J=6.6 Hz), 1.10-1.38 (6H, complex), 1.42-1.77 (2H, complex), 2.48 (1H, dd, J=16.5 and 4.6 Hz), 2.78 (1H, dd, J=16.5 and 9.2 Hz), 2.88 (1H, m), 5.14 (2H, s), 7.23-7.48 (5H, complex), 7.60-9.50 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 1735 (s), 1712 (s)

High resolution MS spectrum [M]⁺ =278.1527 (C₁₆ H₂₂ O₄); Calcd. value: 278.1518 [α]_(D) ²⁶ =+2.4° (c=0.99, EtOH)

Referential Example 8 2,2,2-Trichloroethyl 3-(R)-benzyloxycarbonyloctanoate

Oxalyl chloride (2 ml) was added, under an atmosphere of nitrogen, to a solution of 3-(R)-benzyloxycarbonyloctanoic acid (671 mg), prepared in Referential Example 7, in benzene (10 ml), and the resulting mixture was stirred at 60° C. for 2 hours. Benzene was added thereto, followed by concentration under reduced pressure. After distilling off unreacted oxalyl chloride, the residue was dissolved in tetrahydrofuran (13 ml) under an atmosphere of nitrogen and the solution was cooled to -15° C., followed by the addition of trichloroethanol (1.7 ml) and pyridine (0.23 ml) in turn. The reaction mixture was stirred at -15° C. for 3.3. hours and then poured into 0.5N hydrochloric acid, after which the aqueous mixture was extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography through silica gel, using a 15:1 mixture of hexane and ethyl acetate as an eluent, to give the desired compound (793 mg) as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.85 (3H, t, J=6.6 Hz), 1.12-1.39 (6H, complex), 1.47-1.79 (2H, complex), 2.60 (1H, dd, J=15.5 and 3.3 Hz), 2.89 (1H, dd, J=15.5 and 9.2 Hz), 4.64 (1H, d, J=11.9 Hz), 4.72 (1H, d, J=11.9 Hz), 5.10 (1H, d, J=11.9 Hz), 5.18 (1H, d, J=11.9 Hz), 7.23-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 1757 (s), 1736 (s)

High resolution MS spectrum [M+H]⁺ =409.0734 (C₁₈ H₂₄ O₄ Cl ₃); Calcd. value: 409.0740 [α]_(D) ²⁶ =-1.0° (c=6.0, CHCl₃)

Referential Example 9 2-(R)-(2,2,2-Trichloroethoxycarbonyl)methylheptanoic acid

2,2,2-Trichloroethyl 3-(R)-benzyloxycarbonyloctanoate (924 mg), prepared in Referential Example 8, was dissolved in methanol (8 ml) and was catalytically reduced by stirring the solution for 2 hours under an atmosphere of hydrogen in the presence of 10% palladium on charcoal (52 mg). After completion of the reaction, the catalyst was removed by celite filtration and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography through silica gel using a 30:1 mixture of chloroform and methanol as an eluent, to give the desired compound (678 mg) as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.89 (3H,, t, J=6.5. Hz), 1.18-1.47 (6H, complex), 1.47-1.82 (2H, complex), 2.61 (1H, dd, J=15.2 and 2.9 Hz), 2.88 (1H, dd, J=15.2 and 9.3 Hz), 2.94 (1H, m), 4.72 (1H, d, J=12.0 Hz), 4.79 (1H, d, J=12.0)

IR absorption spectrum (liquid film) cm⁻¹ : 1758 (s), 1709 (s)

High resolution spectrum [M+H]⁺ =319.0261 (C₁₁ H₁₈ O₄ Cl₃); Calcd. value: 319.0271 [α]_(D) ²⁶ =+11.1° (c=3.96, EtOH)

Referential Example 10 N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-2,2,2-trichloroethoxycarbonyl)methylheptyl]-(S)-piperazic acid t-butyl ester

Under an atmosphere of nitrogen, oxalyl chloride (0.6 ml) was added to a solution of 2-(R)-(2,2,2-trichloroethoxycarbonyl)methylheptanoic acid (573 mg), prepared in Referential Example 9, in benzene (10 ml), and the resulting mixture was stirred at 50° C. for 2 hours. After adding dry benzene, the mixture was concentrated under reduced pressure to remove an excess of oxalyl chloride. The residue (acid chloride) thus obtained was dissolved in tetrahydrofuran (4 ml) and the solution was added dropwise to a solution of (S)-N¹ -benzyloxycarbonylpiperazic acid tert-butyl ester (584 mg) and N-ethylmorpholine (0.37 ml) in tetrahydrofuran (7 ml) after cooling to -15° C. under an atmosphere of nitrogen. The resulting mixture was stirred overnight and, during this time, the temperature of the mixture was allowed to rise gradually to room temperature. The reaction mixture was poured into 0.2N hydrochloric acid and extracted with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over anhydrous sodium sulfate. After distilling off the solvent under reduced pressure, the residue was purified by column chromatography through silica gel, using a 6:1 mixture of hexane and ethyl acetate as an eluent, to give the desired compound (1004 mg) as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.80 (3H, t, J=6.6 Hz), 0.85-2.12 (12H, complex), 1.43 (9H, s), 2.61 (1H, dd, J=17.2 and 3.3 Hz), 2.94 (1H, dd, J=17.2 and 10.0 Hz), 3.13 (1H, m), 3.42 (1H, m), 4.28 (1H, br.d, J=11.3 Hz), 4.61 (1H, d, J=11.9 Hz), 4.77 (1H, d, J=11.9 Hz), 5.13 (1H, d, J=11.9 Hz), 5.21 (1H, d, J=11.9 Hz), 5.27 (1H, dd, J=4.6 and 3.9 Hz), 7.22-7.41 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 1739 (s), 1676 (s)

High resolution MS spectrum [M]⁺ =620.1799 (C₂₈ H₃₉ N₂ O₇ Cl₃); Calcd. value: 620.1822 [α]_(D) ²⁶ =-7.5° (c=2.04, CHCl₃)

Referential Example 11 N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxoheptyl]-(S)-piperazic acid t-butyl ester

A 1N aqueous solution of ammonium acetate (2.5 ml) and zinc (1.93 g) were added to a solution of N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methylheptyl]-(S)-piperazic acid tert-butyl ester (920 mg), prepared in Referential Example 10, in tetrahydrofuran, and the resulting mixture was vigorously stirred at room temperature for 4 hours. After completion of the reaction, zinc was filtered off and the filtrate was poured into a 5% aqueous solution of potassium hydrogensulfate followed by extraction with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over anhydrous sodium sulfate. After distilling off the solvent, the residue was purified by column chromatography through silica gel, using a 20:1 mixture of chloroform and methanol as an eluent, to give the desired compound (632 mg) as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.80 (3H, t, J=6.3 Hz), 0.85-2.12 (12H, complex), 1.43 (9H, s), 2.48 (1H, dd, J=17.2 and 4.0 Hz), 2.82 (1H, dd, J=17.2 and 11.1 Hz), 3.09 (1H, m), 3.42 (1H, m), 4.25 (1H, m), 5.12 (1H, d, J=11.9 Hz), 5.21 (1H, d, J=11.9 Hz), 5.27 (1H, dd, J=4.6 and 4.0 Hz), 7.19-7.41 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 1737 (s), 1714 (s), 1675 (s)

High resolution MS spectrum [M+H]⁺ =491.2724 (C₂₆ H₃₉ N₂ O₇); Calcd. value: 491.2756 [α]_(D) ²⁶ =-23.1° (c=1.03 EtOH)

Referential Example 12 N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid tert-butyl ester

N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxoheptyl]-(S)-piperazic acid tert-Butyl ester (598 mg), prepared in Referential Example 11, was dissolved in 20 ml of a 3:1 mixture of tetrahydrofuran and DMF. The solution was cooled to -15° C., O-benzylhydroxylamine (202 mg), diethyl cyanophosphonate (DEPC, 0.28 ml) and triethylamine (0.34 ml) were successively added thereto, and the resulting mixture was stirred for 2.1 hours. The reaction mixture was poured into 1N hydrochloric acid and extracted with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over anhydrous sodium sulfate. After the solvent had been distilled off under reduced pressure, the residue was purified by column chromatography through silica gel, using a 60:1 mixture of chloroform and methanol as an eluent, to give the desired compound (452 mg) and unreacted starting compound (piperazic acid ester, 164 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.79 (3H, t, J=6.8 Hz), 0.82-2.10 (12H, complex), 1.42 (9H, s), 2.10-2.46 (2H, complex), 3.19 (1H, m), 3.42 (1H, m), 4.24 (1H, br.d, J=11.7 Hz), 4.82 (1H, d, J=11.2 Hz), 4.89 (1H, d, J=11.2 Hz), 5.12 (1H, d, J=12.2 Hz), 5.20 (1H, d, J=12.2 Hz), 5.26 (1H, t, J=3.9 Hz), 7.20-7.48 (10H, complex), 7.99 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3426 (m), 1734 (s), 1674 (s)

High resolution MS spectrum [M+H]⁺ =596.3328 (C₃₃ H₄₆ N₃ O₇); Calcd. value: 596.3335 [α]_(D) ²⁶ =-30.7° (c=1.03, CHCl₃)

Referential Example 13 N¹ -Benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid

Trifluoroacetic acid (1.5 ml) was added to a solution of N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid tert-butyl ester (421 mg), prepared in Referential Example 12, in dichloromethane, and the resulting mixture was stirred at room temperature for 2.6 hours. After completion of the reaction, toluene was added to the reaction mixture and the mixture was concentrated under reduced pressure. The residue was purified by column chromatography through silica gel, using a 15:1 mixture of chloroform and methanol as an eluent, to give the desired compound (351 mg) as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.50-3.40 (17H, complex), 4.11 (1H, m), 4.81 (1H, d, J=11.2 Hz), 4.88 (1H, d, J=11.2 Hz), 4.99-5.38 (3H, complex), 7.05-7.55 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3224 (m), 1719 (s), 1672 (s)

Referential Example 14 N-Benzyloxycarbonyl-L-valine (N-methyl-N-methoxy)amide

Following the procedure described in Referential Example 1, but using N-benzyloxycarbonyl-L-valine (2.11 g) as a starting compound, the desired compound (2.21 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.92 (3H, d, J=6.6 Hz), 0.97 (3H, d, J=6.6 Hz), 2.01 (1H, m), 3.21 (3H, s), 3.78 (3H, s), 4.63 (1H, dd, J=9.2 and 7.3 Hz), 5.06 (1H, d, J=12.5 Hz), 5.13 (1H, d, J=12.5 Hz), 5.42 (1H, br.d, J=9.2 Hz), 7.24-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3310 (m), 2975 (m), 1720 (s), 1655 (s) [α]_(D) ²⁶ =+4.4° (c=1.00, CHCl₃)

Referential Example 15 4-(S)-Benzyloxycarbonylamino-5-methyl-3-oxohexane

Following the procedure described in Referential Example 2, but using N-benzyloxycarbonyl-L-valine (N-methyl-N-methoxy)amide (2.21 g) as a starting compound, the desired compound (0.882 g) having a m.p. of 49°-50° C. was obtained after recrystallization from hexane.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.77 (3H, 6., J=6.6 Hz), 1.00 (3H, d, J=6.6 Hz), 1.06 (3H, t, J=7.3 Hz), 2.18 (1H, m), 2.50 (2H, m), 4.36 (1H, dd, J=8.6 and 4.0 Hz), 5.06 (1H, d, J=12.5 Hz), 5.11 (1H, d, J=12.5 Hz), 5.53 (1H, br.d, J=8.6 Hz), 7.23-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3342 (m), 2967 (m), 1712 (s)

High resolution MS spectrum [M+H]⁺ =264.1609 (C₁₅ H₂₂ NO₃); Calcd. value: 264.1600 [α]_(D) ²⁶ =+74.9° (c=0.99, CHCl₃)

Referential Example 16 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (4S)-5-methyl-3-oxohexan-4-ylamide

4-(S)-Benzyloxycarbonylamino-5-methyl-3-oxohexane (61 mg), prepared in Referential Example 15, was dissolved in tetrahydrofuran (2.0 ml) and catalytically reduced by stirring for 40 minutes under an atmosphere of hydrogen in the presence of 10% palladium on charcoal (6 mg). After the catalyst had been filtered off, the filtrate was concentrated under reduced pressure to yield 4-(S)-amino-5-methyl-3-oxohexane as a crude product.

N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (46 mg), prepared in Referential Example 13, was dissolved in 6.4 ml of a 5:3 mixture of tetrahydrofuran and dimethylformamide. The solution was cooled to 0° C. and DEPC (0.06 ml) and a solution of crude 4-(S)-amino-5-methyl-3-oxohexane prepared above in tetrahydrofuran (4.0 ml) were added thereto, and the resulting mixture was stirred for 4.7 hours. After completion of the reaction, the reaction mixture was poured into water and extracted with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over anhydrous sodium sulfate. After the solvent had been distilled off under reduced pressure, the residue was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 2 mm thick), using a 10:1 mixture of chloroform and methanol as a developing solvent, to give the desired compound (75 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.66-0.93 (9H, complex), 1.03 (3H, t, J=7.4 Hz), 1.08-2.60 (17H, complex), 3.13 (1H, m), 3.73 (1H, m), 4.12 (1H, m), 4.40 (1H, br.dd, J=7.8 and 5.6 Hz), 4.82 (1H, d, J=12.6 Hz), 4.89 (1H, d, J=12.6 Hz), 4.95 (1H, m), 5.18 (1H, d, J=12.3 Hz), 5.25 (1H, d, J=12.3 Hz), 7.24-7.38 (10H, complex), 7.95-8.22 (2H, complex)

Referential Example 17 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (4S,5S)-5-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Referential Example 16, but using (4S,5S)-4-benzyloxycarbonylamino-5-methyl-3-oxoheptane (83 mg), prepared in Referential Example 2, and N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (115 mg), the desired compound (85 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.66-2.12 (23H, complex), 1.02 (3H, t, J=7.3 Hz), 1.34 (3H, t, J=7.3 Hz), 2.28 (1H, m), 2.46 (2H, br.q, J=7.3 Hz), 3.12 (1H, m), 4.11 (1H, m), 4.21 (1H, t, J=7.0 Hz), 4.82 (1H, d, J=12.2 Hz), 4.87 (1H, d, J=12.2 Hz), 4.92 (1H, m), 5.17 (1H, d, J=11.7 Hz), 5.25 (1H, d, J=11.7 Hz), 7.24-7.48 (10H, complex), 8.12 (1H, m), 8.27 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3303 (m), 1714 (m), 1667 (s), 1626 (s), 1530 (m)

High resolution MS spectrum [M+H]⁺ =665.3897 (C₃₇ H₅₃ N₄ O₇); Calcd. value: 665.3913

Referential Example 18 N-Benzyloxycarbonyl-D-isoleucine (N-methyl-N-methoxy)-amide

Following the procedure described in Referential Example 1, but using N-benzyloxycarbonyl-D-isoleucine (1.65 g) as a starting material, the desired compound (1.56 g) was obtained as colorless crystals having a m.p. of 64°-66° C., after recrystallization from aqueous methanol.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=7.3 Hz), 0.93 (3H, d, J=6.8 Hz), 1.12 (1H, m), 1.57 (1H, m), 1.73 (1H, m), 3.22 (3H, s), 3.79 (3H, s), 4.67 (1H, dd, J=9.8 and 8.1 Hz), 5.06 (1H, d, J=12.5 Hz), 5.13 (1H, d, J=12.5 Hz), 5.35 (1H, br.d, J=9.8 Hz), 7.23-7.41 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3306 (m), 2965 (m), 1719 (s), 1654 (s)

High resolution MS spectrum [M+H]⁺ =309.1804 (C₁₆ H₂₅ N₂ O₄); Calcd. value: 309.1813 [α]_(D) ²⁶ =+4.54° (c=2.05, CHCl₃)

Referential Example 19 (4R,5R)-4-Benzyloxycarbonylamino-5-methyl-3-oxoheptane

Following the procedure described in Referential Example 2, but using N-benzyloxycarbonyl-D-isoleucine (N-methyl-N-methoxy)amide as a starting compound, there was obtained the desired compound (0.675 g) having a m.p. of 57°-58° C. after recrystallization from aqueous methanol.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=7.3 Hz), 0.98 (3H, d, J=6.8 Hz), 1.04 (1H, m), 1.08 (3H, t, J=7.3 Hz), 1.27 (1H, m), 1.90 (1H, m), 2.52 (2H, m), 4.36 (1H, dd, J=8.3 and 4.6 Hz), 5.09 (2H, s), 5.36 (1H, br.d, J=8.3 Hz), 7.24-7.40 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3270 (w), 2966 (M), 1710 (s)

High resolution. MS spectrum [M+H]⁺ =278.1750 (C₁₆ H₂₄ NO₃); Calcd. value: 278.1756 [α]_(D) ²⁶ =-72.2° (c=1.0, CHCl₃)

Referential Example 20 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (4R,5R)-5-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Referential Example 16, but using (4R,5R)-4-benzyloxycarbonylamino-5-methyl-3-oxoheptane (82 mg), prepared in Referential Example 19, and N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (95 mg), prepared in. Referential Example 13, as starting materials, the desired compound (42 mg) was obtained, having a small amount of impurities but which was able to be used in the following reaction (Referential Example 109) without further purification.

Referential Example 21 N-Benzyloxycarbonyl-L-leucine (N-methyl-N-methoxy)amide

Following the procedure described in Referential Example 1, but using N-benzyloxycarbonyl-L-leucine (2.45 g) as a starting material, the desired compound was obtained (2.61 g).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.93 (3H, d, J=6.6 Hz), 0.97 (3H, d, J=6.6 Hz), 1.47 (2H, t, J=6.6 Hz), 1.72 (1H, m), 3.20 (3H, s), 3.79 (3H, s), 4.79 (1H, m), 5.06 (1H, d, J=12.5 Hz), 5.12 (1H, d, J=12.5 Hz), 5.37 (1H, d, J=9.2 Hz), 7.23-7.41 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3306 (m), 2958 (m), 1720 (s), 1660 (s)

High resolution MS spectrum [M]⁺ =308.1742 (C₁₆ H₂₄ N₂ O₄); Calcd. value: 308.1736 [α]_(D) ²⁶ =-8.4° (c=1.01, CHCl₃)

Referential Example 22 4-(S)-Benzyloxycarbonylamino-6-methyl-3-oxoheptane

Following the procedure described in Referential Example 2, but using N-benzyloxycarbonyl-L-leucine (N=methyl-N-methoxy)amide (2.49 g) as a starting material, the desired compound was obtained (1.36 g).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.92 (3H, d, J=6.6. Hz), 0.98 (3H, d, J=6.6 Hz), 1.08 (3H, t, J=7.3 Hz), 1.37 (1H, ddd, J=14.3, 9.5 and 5.0 Hz), 1.55 (1H, ddd, J=14.3, 9.2 and 4.1 Hz), 1.71 (1H, m), 2.54 (2H, m), 4.42 (1H, m), 5.09 (2H, s), 5.37 (1H, br.d, J=7.3 Hz), 7.24-7.45. (5H, complex)

IR absorption spectrum (film) cm⁻¹ : 3337 (m), 2959 (m), 1713 (s)

High resolution MS spectrum [M-C₂ H₅ --CO]⁺ =220.1358 (C₁₃ H₁₈ NO₂); Calcd. value: 220.1338 [α]_(D) ²⁶ =+32.9° (c=0.99, CHCl₃)

Referential Example 23 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (4S)-6-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Referential Example 16, but using 4-(S)-benzyloxycarbonylamino-6-methyl-3-oxoheptane (57 mg), prepared in Referential Example 22 and N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (42 mg), prepared in Referential Example 13, as starting materials, the desired compound (25 mg) was obtained, having a small amount of impurities but which was able to be used in the following reaction (Example 2) without further purification.

Referential Example 24 2-(S)-[N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1) methyl-1-oxoheptyl]-(S)-piperazyl]aminoisovaleric acid N-methyl-N-methoxyamide

Following the procedure described in Referential Example 16, but using N-benzyloxycarbonyl-L-valine (N-methyl-N-methoxy)amide (96 mg), prepared in Referential Example 14, and N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (48 mg), prepared in Referential Example 13, as starting materials, the desired compound (40 mg) was obtained, having a small amount of impurities but which was able to be used in the following reaction (Example 3) without further purification.

Referential Example 25 Methyl 2-(S)-[N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazyl]aminoisovalerate

N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (41 mg), prepared in Referential Example 13, was dissolved in 3.6 ml of a 3:1 mixture of tetrahydrofuran and dimethylformamide. The solution was cooled to 0° C. and, under an atmosphere of nitrogen, L-valine methyl ester hydrochloride (39 mg), triethylamine (0.025 ml) and DEPC (0.04 ml) were successively added thereto, the resulting mixture was stirred overnight and, during this time, the temperature was allowed to rise gradually to room temperature. After completion of the reaction, the reaction mixture was poured into a 5% aqueous solution of sodium hydrogensulfate and extracted with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over anhydrous sodium sulfate. After the solvent had been distilled off under reduced pressure, the residue was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 15:1 mixture of chloroform and methanol as a developing solvent, to give the desired compound (49 mg) containing a small amount of impurities, but which was able to be used in the following reaction (Example 4) without further purification.

Referential Example 26 tert-Butyl 2-(S)-[N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazinyl]aminoisovalerate

Following the procedure described in Referential Example 25, but using valine tert-butyl ester hydrochloride (45 mg) and N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (37 mg), prepared in Referential Example 13, as starting materials, the desired compound (42 mg) was obtained, having a small amount of impurities, but which was able to be used in the following reaction (Example 5) without further purification.

Referential Example 27 N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonylmethyl)heptyl]-(R)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 10, but using 2-(R)-(2,2,2-trichloroethoxycarbonylmethyl)heptanoic acid (246 mg) and (R)-N¹ -benzyloxycarbonylpiperazic acid tert-butyl ester (246 mg) as starting materials, the desired compound (267 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.85 (3H, t, J=6.6 Hz), 0.97-2.14 (12H, complex), 1.43 (3H, s), 2.50 (1H, dd, J=17.5 and 4.9 Hz), 2.95 (1H, dd, J=17.5 and 9.9 Hz), 2.97 (1H, m), 3.28 (1H, m), 4.40 (1H, m), 4.58 (1H, d, J=12.5 Hz), 4.85 (1H, d, J=12.5 Hz), 5.10 (1H, d, J=12.5 Hz), 5.21 (1H, d, J=12.5 Hz), 5.29 (1H, br.d, J=4.3 Hz), 7.22-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2931 (m), 1735 (s), 1677 (s)

High resolution MS spectrum [M]⁺ =620.1833 (C₂₈ H₃₉ N₂ O₇ ³⁵ Cl₃); Calcd. value: 620.1823

Referential Example 28 N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxoheptyl]-(R)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 11, but using N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonylmethyl)heptyl]-(R)-piperazic acid tert-butyl ester (694 mg), prepared in Referential Example 27, there was obtained the desired compound (533 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.84 (3H, t, J=6.6 Hz), 0.94-2.17 (12H, complex), 1.42 (9H, s), 2.37 (1H, m), 2.80-3.09 (2H, complex), 3.18 (1H, m), 4.39 (1H, m), 5.00-5.36 (3H, complex), 7.18-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3190 (w), 2932 (s), 1735 (s), 1679 (s)

High resolution MS spectrum [M+H-H₂ O]⁺ =473.2672 (C₂₆ H₃₇ N₂ O₆); Calcd. value: 473.2652

Referential Example 29 N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 12, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxoheptyl]-(R)-piperazic acid tert-butyl ester (517 mg), prepared in Referential Example 28, and O-benzylhydroxylamine (203 mg) as starting materials, the desired compound (421 mg), was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 0.93-2.38 (14H, complex), 1.42 (9H, s), 2.81-3.32 (2H, complex), 4.32 (1H, m), 4.75-4.95 (2H, complex), 5.10 (1H, d, J=11.9 Hz), 5.20 (1H, d, J=11.9 Hz), 5.27 (1H, m), 7.22-7.46 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3252 (m), 2931 (s), 1735 (s), 1675 (s)

High resolution MS spectrum [M+H]⁺ =596.3327 (C₃₃ H₄₆ N₃ O₇); Calcd. value: 596.3335 [α]_(D) ²⁶ =+37.1° (c=1.00, EtOH)

Referential Example 30 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid

Following the procedure described in Referential Example 13, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-(2-benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid tert-butyl ester (56 mg), prepared in Referential Example 29, the desired compound (43 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.83 (3H, t, J=5.8 Hz), 0.97-2.20 (15H, complex), 3.14 (1H, m), 4.24 (1H, m), 4.70-4.95 (2H, br.s), 5.02-5.33 (3H, complex), 7.18-7.48 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3230 (w), 2940 (m), 1720 (s), 1655 (s) [α]_(D) ²⁶ =+21.4° (c=1.0, EtOH)

Referential Example 31 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid (4S,5S)-5-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Referential Example 16, but using (4S,5S)-4-benzyloxycarbonylamino-5-methyl-3-oxoheptane (61 mg), prepared in Referential Example 2, and N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid (74 mg), prepared in Referential Example 30, as starting materials, the desired compound (67 mg) was obtained, having a small amount of impurities but which was able to be used in the following reaction (Referential Example 110) without further purification.

Referential Example 32 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid (4R,5R)-5-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Referential Example 16, but using (4R,5R)-4-benzyloxycarbonylamino-5-methyl-3-oxoheptane (55 mg), prepared in Referential Example 19, and N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid (72 mg), prepared in Referential Example 30, the desired compound (54 mg) was obtained, having a small amount of impurities but which was able to be used in the following reaction (Referential Example 111) without further purification.

Referential Example 33 N¹ -benzyloxycarbonyl-N² -[3-(2,2,2-trichloroethoxycarbonyl)propionyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 10, but using mono-2,2,2-trichloroethyl succinate (0,253 g) and (S)-N¹ -benzyloxycarbonylpiperazic acid tert-butyl ester (0.323 g) as starting materials, the desired compound (0.582 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.41 (9H, s), 1.48-2.13 (4H, complex), 2.38-3.21 (5H, complex), 4.66 (1H, d, J=12.0 Hz), 4.79 (1H, d, J=12.0 Hz), 4.92-5.42 (3H, complex), 7.25-7.40 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2977 (m), 1731 (s), 1683 (s)

High resolution MS spectrum [M]⁺ =550.1028 (C₂₃ H₂₉ N₂ O₇ ³⁵ Cl₃); Calcd. value: 550.1040 [α]_(D) ²⁶ =-21.1° (c=1.01, CHCl₃)

Referential Example 34 N¹ -benzyloxycarbonyl-N² -(3-carboxypropionyl)-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 11, but using N¹ -benzyloxycarbonyl-N² -[3-(2,2,2-trichloroethoxycarbonyl)propionyl]-(S)-piperazic acid tert-butyl ester (0.543 g), prepared in Referential Example 33, as starting materials, the desired compound (0,381 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.4 (9H, s), 1.55 (1H, m), 1.78 (1H, m), 1.88-2.11 (2H, complex), 2.33-2.89 (5H, complex), 3.01 (1H, m), 4.91-5.44 (3H, complex), 7.22-7.45 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3195 (m), 2978 (m), 1730 (s), 1683 (s)

High resolution MS spectrum [M+H]⁺ =421.2019 (C₂₁ H₂₉ N₂ O₇); Calcd. value: 421.1975

Referential Example 35 N¹ -benzyloxycarbonyl-N² -(3-benzyloxyaminocarbonylpropionyl)-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 12, but using N¹ -benzyloxycarbonyl-N² -(3-carboxypropionyl)-(S)-piperazic acid tert-butyl ester (350 mg), prepared in Referential Example 34, and O-benzylhydroxylamine (161 mg) as starting materials, the desired compound (458 mg) was obtained, having a small amount of impurities but which was able to be used in the following reactions (Examples 7 and Referential Example 36) without further purification.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.42 (9H, s), 1.50-2.12 (4H, complex), 2.48 (2H, m), 2.48 (2H, m), 2.80 (2H, m), 3.09 (2H, m), 4.87 (2H, s), 4.96-5.41 (3H, complex), 7.21-7.47 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3240 (m), 2990 (m), 1725 (s), 1675 (s)

High resolution MS spectrum [M+H]⁺ =526.2546 (C₂₈ H₃₆ N₃ O₇); Calcd. value: 526.2552

Referential Example 36 N¹ -Benzyloxycarbonyl-N² -(3-benzyloxyaminocarbonylpropionyl)-(S)-piperazic acid

Following the procedure described in Referential Example 13, but using N¹ -benzyloxycarbonyl-N² -(3-carboxypropionyl)-(S)-piperazic acid tert-butyl ester (369 mg), prepared in Referential Example 34, as a starting material, there was obtained the desired compound (299 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.60 (1H, m), 1.69-2.60 (6H, complex), 2.68-2.98 (2H, complex), 3.31 (1H, m), 4.87 (2H, br.s), 3.10-3.32 (3H, complex), 7.18-7.47 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3250 (m), 1720 (s), 1670 (s) [α]_(D) ²⁶ =-12.8° (c=2.01 EtOH)

Referential Example 37 N¹ -Benzyloxycarbonyl-N² -(3-benzyloxyaminocarbonylpropionyl)-(S)-piperazic acid (4S,5S)-5-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Referential Example 16, but using (4S,5S)-4-benzyloxycarbonylamino-5-methyl-3-oxoheptane (110 mg) and N¹ -benzyloxycarbonyl-N² -(3-benzyloxyaminocarbonylpropionyl)-(S)-piperazic acid (159 mg), prepared in Referential Example 36, the desired compound (128 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, d, J=6.6. Hz), 1.01-2.00 (7H, complex), 1.02 (3H, t, J=7.2 Hz), 1.38 (3H, t, J=7.2 Hz), 2.02-2.69 (4H, complex), 2.48 (2H, q, J=7.2 Hz), 2.81 (1H, m), 3.32 (1H, m), 4.48 (1H, dd, J=7.3 and 6.6 Hz), 4.89 (2H, s), 5.10 (1H, m), 5.15 (1H, d, J=12.5 Hz), 5.25 (1H, d, J=12.5 Hz), 7.25-7.50 (10H, complex), 7.98 (1H, br.d, J=8.0 Hz), 8.65 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3320 (m), 2975 (m), 1720 (s), 1700 (s), 1675 (s)

High resolution MS spectrum [M+H]⁺ =595.3134 (C₃₂ H₄₃ N₄ O₇); Calcd. value: 595.3131

Referential Example 38 4-(S)-Isopropyl-3-(1-oxo-4-methylpentyl)-2-oxazolidinone

Following the procedure described in Referential Example 3, but using 4-(S)-isopropyl-2-oxazolidone (3.46 g) and isocaproyl chloride (3.98 g) as starting materials, the desired compound (5.21 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.83-1.00 (12H, complex), 1.45-1.72 (3H, complex), 2.37 (1H, dhep, J=4.0 and 7.9 Hz), 2.78-3.08 (2H, complex), 4.21 (1H, dd, J=7.9 and 4.0 Hz), 4.27 (1H, t, J=7.9 Hz), 4.44 (1H, dt, J=7.9 and 4.0 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 2970 (s), 1780 (s), 1700 (s)

High resolution. MS spectrum [M]⁺ =227.1522 (C₁₂ H₂₁ NO₃); Calcd. value: 227.1521 [α]_(D) ²⁶ =+76.3° (c=1.00, CHCl₃)

Referential Example 39 4-(S)-isopropyl-3-[2-(R)-tert-butoxycarbonylmethyl-1-oxo-4-methylpentyl]-2-oxazolidinone

Following the procedure described in Referential Example 4, but using 4-(S)-isopropyl-3-(1-oxo-4-methylpentyl)-2-oxazolidinone (5.17 g), prepared in Referential Example 38, and tert-butyl bromoacetate (18.5 ml) as starting materials, the desired compound (6.16 g), having a m.p. of 143°-144° C. after recrystallization from aqueous methanol, was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86-0.97 (12H, complex), 1.30 (1H, m), 1.42 (9H, s), 1.45-1.66 (3H, complex), 2.38 (1H, m), 2.44 (1H, dd, J=16.5 and 5.3 Hz), 2.68 (1H, dd, J=16.5 and 9.9 Hz), 4.18-4.31 (2H, complex), 4.42 (1H, dt, J=7.9 and 4.0 Hz)

IR absorption spectrum (film) cm⁻¹ : 2960 (m), 1764 (s), 1732 (s), 1700 (s)

High resolution MS spectrum [M+H]⁺ =342.2270 (C₁₈ H₃₂ NO₅); Calcd. value: 342.2280 [α]_(D) ²⁶ =+41.6° (c=1.00, CHCl₃)

Referential Example 40 2-(R)-(tert-Butoxycarbonylmethyl)-4-methylpentanoic acid

Following the procedure described in Referential Example 5, but using 4-(S)-isopropyl-3-[2-(R)-tert-butoxycarbonylmethyl-1-oxo-4-methylpentyl]-2-oxazolidinone (6.14 g), prepared in Referential Example 39, as a starting material, the desired compound (3.99 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.91 (3H, d, J=6.6 Hz), 0.94 (3H, d, J=6.6 Hz), 1.29 (1H, m), 1.43 (9H, s), 1.53-1.77 (2H, complex), 2.37 (1H, dd, J=16.3 and 5.3 Hz), 2.59 (1H, dd, J=16.3 and 9.2 Hz), 2.85 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 2960 (s), 1731 (s), 1710 (s) [α]_(D) ²⁶ =+14.1° (c=1.00, EtOH)

Referential Example 41 tert -Butyl 3-(R)-Benzyloxycarbonyl-5-methylhexanoate

Following the procedure described in Referential Example 6, but using 2-(R)-(tert-butoxycarbonylmethyl)-4-methylpentanoic acid (2.88 g), prepared in Referential Example 40, as a starting material, the desired compound (2.17 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, d, J=6.6 Hz), 0.91 (3H, d, J=6.6 Hz), 1.29 (1H, m), 1.41 (9H, s), 1.49-1.68 (2H, complex), 2.35 (1H, dd, J=16.6 and 5.9 Hz), 2.62 (1H, dd, J=16.6 and 8.4 Hz), 2.90 (1H, m), 5.10 (1H, d, J=12.5 Hz), 5.16 (1H, d, J=12.5 Hz), 7.25-7.40 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2959 (m), 1732 (s)

High resolution MS spectrum [M+H]⁺ =321.2051 (C₁₉ H₂₉ O₄); Calcd. value: 321.2066 [α]_(D) ²⁶ =+2.4° (c=4.96, CHCl₃)

Referential Example 42 3-(R)-Benzyloxycarbonyl-5-methylhexanoic acid

Following the procedure described in Referential Example 7, but using tert-butyl 3-(R)-benzyloxycarbonyl-5-methylhexanoate (2.10 g), prepared in Referential Example 41, as a starting material, the desired compound (1.73 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, d, J=6.6 Hz), 0.92 (3H, d, J=6.6 Hz), 1.31 (1H, m), 1.48-1.69 (2H, complex), 2.48 (1H, dd, J=17.0 and 4.6 Hz), 2.76 (1H, dd, J=17.0 and 9.6 Hz), 2.94 (1H, m), 5.12 (1H, d, J=12.9 Hz), 5.16 (1H, d, J=12.9 Hz), 7.23-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2959 (m), 1736 (s), 1713 (s)

High resolution MS spectrum [M]⁺ =264.1350 (C₁₅ H₂₀ O₄); Calcd. value: 264.1362 [α]_(D) ²⁶ =+6.8° (c=1.00, EtOH)

Referential Example 43 2,2,2-Trichloroethyl 3-(R)-Benzyloxycarbonyl-5-methylhexanoate

Following the procedure described in Referential Example 8, but using 3-(R)-benzyloxycarbonyl-5-methylhexanoic acid (1.70 g), prepared in Referential Example 42, as a starting material, the desired compound (2.20 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, d, J=6.6 Hz), 0.92 (3H, d, J=6.6 Hz), 1.38 (1H, m), 1.48-1.72 (2H, m), 2.67 (1H, m), 2.87 (1H, m), 3.01 (1H, m), 4.64 (1H, d, J=12.9 Hz), 4.74 (1H, d, J=12.9 Hz), 5.11 (1H, d, J=13.2 Hz), 5.16 (1H, d, J=13.2 Hz), 7.23-7.43 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2958 (m), 1758 (s), 1736 (s) [α]_(D) ²⁶ =+1.9° (c=4.03, CHCl₃)

Referential Example 44 2-(R)-(2,2,2-Trichloroethoxycarbonyl)methyl-4-methylpentanoic acid

Following the procedure described in Referential Example 9, but using 2,2,2-trichloroethyl 3-(R)-benzyloxycarbonyl-5-methylhexanoate (1.79 g), prepared in Referential Example 43, as a starting material, the desired compound (1.12 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.93 (3H, d, J=6.6 Hz), 0.96 (3H, d, J=6.6 Hz), 1.40 (1H, m), 1.55-1.79 (2H, complex), 2.61 (1H, dd, J=16.5 and 4.6 Hz), 2.85 (1H, dd, J=16.5 and 9.2 Hz), 2.97 (1H, m), 4.72 (1H, d, J=12.2 Hz), 4.79 (1H, d, J=12.2 Hz)

IR absorption spectrum (film) cm⁻¹ : 2960 (m), 1758 (s), 1710 (s)

High resolution MS spectrum [M+H]⁺ =305.0100 (C₁₀ H₁₆ O₄ ³⁵ Cl₃); Calcd. value: 305.0114 [α]_(D) ²⁶ =+11.4° (c=1.08, EtOH)

Referential Example 45 N¹ -Benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-4-methyl-1-oxopentyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 11, but using N¹ -Benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methyl-4-methylpentyl]-(S)-piperazic acid tert-butyl ester (625 mg), prepared in Referential Example 107, as a starting material, the desired compound (477 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.83 (3H, d, J=6.6 Hz), 0.90 (3H, d, J=6.6 Hz), 1.11-1.62 (4H, complex), 1.43 (9H, s), 1.70-2.12 (3H, complex), 2.52 (1H, dd, J=17.8 and 3.3 Hz), 2.77, (1H, dd, J=17.8 and 10.6 Hz), 3.17 (1H, br.t, J=10.6 Hz), 3.36 (1H, br.t, J=10.9 Hz), 4.24 (1H, m), 5.13 (1H, d, J=12.2 Hz), 5.20 (1H, d, J=12.2 Hz), 5.26 (1H, dd, J=4.6 and 4.0 Hz), 7.22-7.46 (5H, complex)

IR absorption spectrum (film) cm⁻¹ : 3186 (w), 2958 (m), 1736 (s), 1677 (s)

High resolution MS spectrum [M+H]⁺ =477.2612 (C₂₅ H₃₇ N₂ O₇); Calcd. value: 477.2601 [α]_(D) ²⁶ =19.6° (c=1.01, EtOH)

Referential Example 46 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-4-methyl-1-oxopentyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 12, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-4-methyl-1-oxopentyl]-(S)-piperazic acid tert-butyl ester (459 mg), prepared in Referential Example 45, and O-benzylhydroxylamine (234 mg) as starting materials, there was obtained the desired compound (427 mg), which had a small amount of impurities but which was able to be used in the following reaction (Example 11) without further purification.

Referential Example 47 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N-methylamide

Following the procedure described in Referential Example 25, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxycarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (39 mg) and methylamine hydrochloride (17 mg) as starting materials, the desired compound (14 mg) was obtained, having a small amount of impurities but which was able to be used in the following reaction (Referential Example 48) without further purification.

Referential Example 48 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N-methylamide

Following the procedure described in Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxycarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N-methylamide (14 mg), prepared in Referential Example 47, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography silica gel (20×20 cm size, 0.5 mm thick), using a 15:1 mixture of chloroform and methanol as a developing solvent twice, to give the desired compound (5.0 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.00-2.13 (12H, complex), 2.30 (1H, m), 2.52 (1H, br.t, J=12.5 Hz), 2.79 (3H, d, J=4.5 Hz), 2.82 (1H, m), 3.02 (1H, br.d, J=13.2 Hz), 3.85 (1H, m), 4.61 (1H, d, J=11.9 Hz), 5.05 (1H, br.s), 6.59 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3270 (m), 2940 (m), 1655 (s), 1625 (s)

High resolution MS spectrum [M]⁺ =328 [α]_(D) ²⁶ =-8.8° (c=0.45, EtOH)

Referential Example 49 4-(S)-Isopropyl-3-(1-oxodecyl)-2-oxazolidinone

Following the procedure described in Referential Example 3, but using 4-(S)-isopropyl-2-oxazolidone (4.74 g) and decanoyl chloride (7.35 g) as starting materials, the desired compound (9.85 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.8 Hz), 0.87 (3H, d, J=6.6 Hz), 0.92 (3H, d, J=6.6 Hz), 1.17-1.43 (12H, complex), 1.59-1.73 (2H, complex), 2.37 (1H, d.hep, J=3.3 and 6.6 Hz), 2.78-3.05 (2H, complex), 4.20 (1H, dd, J=9.2 and 3.3 Hz), 4.26 (1H, dd, J=9.2 and 7.9 Hz), 4.44 (1H, d, t, J=7.9 and 3.3 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 2927 (s), 1785 (s), 1703 (s)

High resolution spectrum [M]⁺ =283.2144 (C₁₆ H₂₉ NO₃); Calcd. value: 283.2147 [α]_(D) ²⁶ =+61.4° (c=1.00, CHCl₃)

Referential Example 50 4-(S)-Isopropyl-3-[2-(R)-tert-butoxycarbonylmethyl-1-oxodecyl)-2-oxazolidinone

Following the procedure described in Referential Example 4, but using 4-(S)-isopropyl-3-(1-oxodecyl)-2-oxazolidinone (9.85 g), prepared in Referential Example 49, and tert-butyl bromoacetate (25.4 ml) as starting materials, the desired compound (9.17 g) having a m.p. of 58°-59° C. was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.8 Hz), 0.88 (3H, d, J=6.6 Hz), 0.93 (3H, d, J=6.6 Hz), 1.13-1.37 (12H, complex), 1.41 (9H, s), 1.53-1.70 (2H, complex), 2.38 (1H, d.hep, J=3.3 and 6.6 Hz), 2.43 (1H, dd, J=16.6 and 4.7 Hz), 2.74 (1H, dd, J=16.6 and 10.6 Hz), 4.08-4.30 (3H, complex), 4.44 (1H, dt, J=7.3 and 3.3 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 2925 (m), 1764 (s), 1730 (s), 1702 (s)

High resolution MS spectrum [M+H]⁺ =398.2910 (C₂₂ H₄₀ NO₅); Calcd. value: 398.2907 [α]_(D) ²⁶ =+46.6° (c=1.00, CHCl₃)

Referential Example 51 2-(R)-(tert-Butoxycarbonylmethyl)decanoic acid

Following the procedure described in Referential Example 5, but using 4-(S)-isopropyl-3-[2-(R)-tertbutoxycarbonylmethyl-1-oxodecyl]-2-oxazolidinone (9.08 g), prepared in Referential Example 50, as a starting material, the desired compound (6.14 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.18-1.40 (12H, complex), 1.43 (9H, s), 1.51 (1H, m), 1.67 (1H, m), 2.38 (1H, dd, J=16.5 and 5.3 Hz), 2.61 (1H, dd, J=16.5 and 9.2 Hz), 2.80 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 2858 (s), 1733 (s), 1709 (s)

High resolution MS spectrum [M+H]⁺ =287.2213 (C₁₆ H₃₁ O₄); Calcd. value: 287.2223 [α]_(D) ²⁶ =+14.2° (c=1.00, EtOH)

Referential Example 52 tert-Butyl 3-(R)-Benzyloxycarbonylundecanoate

Following the procedure described in Referential Example 6, but using 2-(R)-(tert-butoxycarbonylmethyl)decanoic acid (4.64 g), prepared in Referential Example 51, as a starting material, the desired compound (5.69 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.10-1.34 (12H, complex), 1.41 (9H, s), 1.50 (1H, m), 1.61 (1, m), 2.36 (1H, dd, J=16.2 and 5.3 Hz), 2.64 (1H, dd, J=16.2 and 9.2 Hz), 2.83 (1H, m), 5.09 (1H, d, J=12.5 Hz), 5.17 (1H, d, J=12.5 Hz), 7.28-7.44 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2956 (m), 1733 (s)

High resolution MS spectrum [M+H]⁺ =377.2658 (C₂₃ H₃₇ O₄); Calcd. value: 377.2692 [α]_(D) ²⁶ =+1.1° (c=1.00, CHCl₃)

Referential Example 53 3-(R)-Benzyloxycarbonylundecanoic acid

Following the procedure described in Referential Example 7, but using tert-butyl 3-(R)-benzyloxycarbonylundecanoate (5.62 g), prepared in Referential Example 52, as a starting material, the desired compound (4.07 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6. Hz), 1.10-1.38 (12H, complex), 1.44-1.73 (2H, complex), 2.49 (1H, dd, J=16.2 and 4.3 Hz), 2.79 (1H, dd, J=16.2 and 9.6 Hz), 2.87 (1H, m), 5.12 (1H, d, J=13.2 Hz), 5.17 (1H, d, J=13.2 Hz), 7.13-7.43 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2927 (s), 1737 (s), 1713 (s)

High resolution MS spectrum [M]⁺ =320.1989 (C₁₉ H₂₈ O₄); Calcd. value: 320.1987 [α]_(D) ²⁶ =+3.3° (c=1.00, EtOH)

Referential Example 54 2,2,2-Trichloroethyl 3-(R)-Benzyloxycarbonylundecanoate

Following the procedure described in Referential Example 8, but using 3-(R)-benzyloxycarbonylundecanoic acid (3.65 g), prepared in Referential Example 53, as a starting material, the desired compound (3.18 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.14-1.38 (12H, complex), 1.48-1.79 (2H, complex), 2.60 (1H, dd, J=15.2 and 4.0 Hz), 2.90 (1H, dd, J=15.2 and 9.2 Hz), 2.96 (1H, m), 4.65 (1H, d, J=12.9 Hz), 4.73 (1H, d, J=12.9 Hz), 5.10 (1H, d, J=12.2 Hz), 5.18 (1H, d, J=12.2 Hz), 7.28-7.43 (5H, complex)

IR absorption spectrum (film) cm⁻¹ : 2927 (s), 1737 (s), 1713 (s)

High resolution MS spectrum [M]⁺ =452.1099 (C₂₁ H₂₉ O₄ ³⁵ Cl₂ ³⁷ Cl); Calcd. value: 452.1102 [α]_(D) ²⁶ =0.44° (c=5.00, CHCl₃)

Referential Example 55 2-(R)-(2,2,2-Trichloroethoxycarbonyl)methyldecanoic acid

Following the procedure described in Referential Example 9, but using 2,2,2-trichloroethyl 3-(R)-benzyloxycarbonylundecanoate (3.08 g), prepared in Referential Example 54, as a starting material, the desired compound (1.87 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.17-1.46 (12H, complex), 1.51-1.82 (2H, complex), 2.61 (1H, dd, J=15.1 and 3.0 Hz), 2.81-3.10 (2H, complex), 4.72 (1H, d, J=12.2 Hz), 4.79 (1H, d, J=12.2 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 2928 (s), 1759 (s), 1710 (s)

High resolution MS spectrum [M+H]⁺ =361.0736 (C₁₄ H₂₄ O₄ ³⁵ Cl₃); Calcd. value: 361.0740 [α]_(D) ²⁶ =+11.5° (c=1.00, EtOH)

Referential Example 56 N¹ -Benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methyldecyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 10, but using 2-(R)-(2,2,2-trichloroethoxycarbonyl)methyldecanoic acid (583 mg), prepared in Referential Example 55, and (S)-N¹ -benzyloxycarbonylpiperazic acid tert-butyl ester (490 mg) as starting materials, the desired compound was obtained (704 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 0.93-2.14 (18H, complex), 1.43 (9H, s), 2.60 (1H, dd, J=17.5 and 3.6 Hz), 2.95 (1H, dd, J=17.5 and 10.9 Hz), 3.14 (1H, m), 3.42 (1H, br.t, J=11.3 Hz), 4.27 (1H, br.d, J=12.5 Hz), 4.61 (1H, d, J=12.2 Hz), 4.78 (1H, d, J=12.2 Hz), 5.14 (1H, d, J=11.9 Hz), 5.21 (1H, d, J=11.9 Hz), 5.27 (1H, t, J=4.3 Hz), 7.23-7.40 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2929 (m), 1740 (s), 1677 (s)

High resolution MS spectrum [M+2H]⁺ =664.2264 (C₃₁ H₄₅ O₇ ³⁵ Cl₃); Calcd. value: 664.2263 [α]_(D) ²⁶ =-6.8° (c=1.00, CHCl₃)

Referential Example 57 N¹ -Benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxydecyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 11 but using N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methyldecyl]-(S)-piperazic acid tert-butyl ester (645 mg), prepared in Referential Example 56, as a starting material, the desired compound was obtained (455 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 0.93-2.11 (18H, complex), 1.43 (9H, s), 2.49 (1H, dd, J=17.2 and 3.3 Hz), 2.82 (1H, dd, J=17.2 and 11.2 Hz), 3.09 (1H, m), 3.32 (1H, dd, J=14.2 and 10.7 Hz), 4.25 (1H, br.d, J=8.5 Hz), 5.13 (1H, d, J=12.5 Hz), 5.20 (1H, d, J=12.5 Hz), 5.27 (1H, d, J=4.3 Hz), 7.20-7.40 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3189 (w), 2929 (m), 1736 (s), 1678 (s)

High resolution MS spectrum [M+H]⁺ =533.3249 (C₂₉ H₄₅ N₂ O₇); Calcd. value: 533.3227 [α]_(D) ²⁶ =-21.3° (c=1.00, EtOH)

Referential Example 58 N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxydecyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 12 but using N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxodecyl]-(S)-piperazic acid tert-butyl ester (437 mg), prepared in Referential Example 57, as a starting material, the desired compound was obtained (490 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 0.92-2.52 (18H, complex), 1.43 (9H, s), 2.10-2.46 (2H, complex), 3.20 (1H, m), 3.41 (1H, m), 4.24 (1H, m), 4.82 (1H, d, J=11.6 Hz), 4.89 (1H, d, J=11.6 Hz), 5.12 (1H, d, J=12.5 Hz), 5.20 (1H, d, J=12.5 Hz), 5.27 (1H, t, J=2.4 Hz), 7.22-7.48 (10H, complex), 8.12 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3255 (w), 2928 (s), 1733 (s), 1675 (s)

High resolution MS spectrum [M+H--tBu]⁺ =581.3104 (C₃₂ H₄₃ N₃ O₇); Calcd. value: 581.3101 [α]_(D) ²⁶ =-34.30° (c=1.00, CHCl₃)

Referential Example 59 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid

Following the procedure described in Referential Example 13, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid tert-butyl ester (433 mg), prepared in Referential Example 58, as a starting material, the desired compound (379 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 0.92-2.59 (20H, complex), 2.51-3.18 (2H, complex), 4.10 (1H, br.d, J=12.5 Hz), 4.83 (1H, t, J=4.5 Hz), 4.93 (1H, d, J=11.2 Hz), 5.00 (1H, d, J=11.2 Hz), 5.01 (1H, d, J=11.9 Hz), 5.18 (1H, d, J=11.9 Hz), 7.14 (1H, br.s), 7.20-7.51 (10H, complex), 12.3 (1H, br.s)

IR absorption spectrum (FT film) cm⁻¹ : 3228 (w), 2927 (s), 1705 (s), 1672 (s), 1604 (s) [α]_(D) ²⁶ =-29.0° (c=1.00, EtOH)

Referential Example 60 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N-methylamide

Following the procedure described in Referential Example 25, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid tert-butyl ester (44 mg), prepared in Referential Example 59, and methylamine hydrochloride (10 mg) as starting materials, the desired compound (15 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 0.98-2.59 (20H, complex), 2.70 (3H, d, J=4.6 Hz), 3.08 (1H, m), 3.67 (1H, m), 4.11 (1H, m), 4.83 (1H, d, J=11.2 Hz), 4.88 (1H, d, J=11.2 Hz), 5.13 (1H, t, J=5.3 Hz), 5.21 (2H, s), 7.25-7.49 (10H, complex), 7.71 (1H, m), 8.18 (1H, m)

IR absorption spectrum (film) cm⁻¹ : 3345 (m), 3225 (m), 2945 (s), 1695 (s), 1670 (s)

Mass spectrum m/z [M-H₂ O]⁺ =576

Referential Example 61 4-(S)-Isopropyl-3-(1-oxooctyl)-2-oxazolidinone

Following the procedure described in Referential Example 3, but using 4-(S)-isopropyl-2-oxazolidinone (5.06 g) and octanoyl chloride (6.67 g), the desired compound (9.39 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, d, J=6.6 Hz), 0.88 (3H, t, overlapped to 0.87 ppm and 0.92 ppm), 0.92 (3H, d, J=6.6 Hz), 1.09-1.42 (8H, complex), 1.50-1.73 (2H, complex), 2.38 (1H, m), 2.70-3.06 (2H, complex), 4.20 (1H, dd, J=9.2 and 3.3 Hz), 4.26 (1H, t, J=9.2 Hz), 4.44 (1H, dt, J=9.2 and 3.3 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 2929 (m), 1784 (s), 1702 (s)

Mass spectrum [M]⁺ =255

High resolution MS spectrum [M]⁺ =255.1832 (C₁₄ H₂₅ O₃ N); Calcd. value: 255.1835 [α]_(D) ²⁶ =+74.9° (c=1.0, CHCl₃)

Referential Example 62 4-(S)-Isopropyl-3-[2-(R)-tert-butoxycarbonylmethyl-1-oxooctyl]-2-oxazolidinone

Following the procedure described in Referential Example 4, but using 4-(S)-isopropyl-3-(1-oxooctyl)-2-oxazolidinone (5.51 g), prepared in Referential Example 61, and tert-butyl bromoacetate (17 ml), the desired compound (4.13 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=7.3 Hz), 0.91 (3H, d, J=6.3 Hz), 0.93 (3H, d, J=6.3 Hz), 1.15-1.50 (9H, complex), 1.41 (9H, s), 1.62 (1H, m), 2.36 (1H, m), 2.42 (1H, dd, J=16.6 and 4.4 Hz), 2.74 (1H, dd, J=16.6 and 10.3 Hz), 4.08-4.30 (3H, complex), 4.43 (1H, dt, J=7.6 and 3.7 Hz)

IR absorption spectrum (liquid film, kBr pellet): 2931 (m), 1764 (s), 1730 (s), 1703 (s)

Mass Spectrum. [M+H]⁺ =370

High resolution MS spectrum [M+H]⁺ =370.2587 (C₂₀ H₃₆ NO₅); Calcd. value: 370.2593

m.p. 50°-51° C. (H₂)--MeOH) [α]_(D) ²⁶ =+51.9° (c=1.0, CHCl₃)

Referential Example 63 tert-Butyl 3-(R)-benzyloxycarbonylnonanoate

4-(S)-Isopropyl-3-[2-(R)-tert-butoxycarbonylmethyl-1-oxooctyl]-2-oxazolidinone (3.98 g), prepared in Referential Example 62 was dissolved in tetrahydrofuran. The solution was cooled to 0° C., and a lithium benzyloxide solution (42 ml), which was prepared from tetrahydrofuran (30 ml), benzyl alcohol (2.4 ml) and a 1.66M solution (9.8 ml) of n-butyllithium in hexane with ice-cooling, was added thereto, and the resulting mixture was stirred at 0° C. for 40 minutes. The reaction mixture was poured into a 5% aqueous solution of potassium hydrogensulfate and extracted with ethyl acetate. The organic extract was successively washed with water and a saturated aqueous solution of sodium chloride, and dried over sodium sulfate. After the solvent had been distilled off under reduced pressure, the residue was purified by column chromatography through silica gel, using a 20:1 mixture of hexane and ethyl acetate as an eluent, to give the desired compound (3.86 g) as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=6.6 Hz), 1.10-1.72 (10H, complex), 1.41 (9H, s), 2.36 (1H, dd, J=16.2 and 5.3 Hz), 2.64 (1H, dd, J=16.2 and 9.2 Hz), 2.83 (1H, m), 5.09 (1H, d, J=12.5 Hz), 5.17 (1H, d, J=12.5 Hz), 7.26-7.41 (5H, complex)

IR absorption. Spectrum (liquid film) cm⁻¹ : 2931 (m), 1732 (s)

Mass spectrum [M+H--tBu]⁺ =292

High resolution MS spectrum [M+H--tBu]⁺ =292.1677 (C₁₇ H₂₄ O₄); Calcd. value: 292.1675 [α]_(D) ²⁶ =+0.38° (c=6.9, CHCl₃)

Referential Example 64 3-(R)-Benzyloxycarbonylnonanoic acid

Following the procedure described in Referential Example 7, but using tert-butyl 3-(R)-benzyloxycarbonylnonanoate (3.86 g), prepared in the Referential Example 63, the desired compound (2.73 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=6.6 Hz), 1.09-1.37 (8H, complex), 1.41-1.78 (2H<complex), 2.49 (1H, dd, J=16.3 and 4.4 Hz), 2.79 (1H, dd, J=6.3 and 9.3 Hz), 2.88 (1H, m), 5.12 (1H, d, J=12.2 Hz), 5.17 (1H, d, J=12.2 Hz), 7.23-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2930 (m), 1736 (s), 1712 (s)

Mass spectrum [M]⁺ =292

High resolution MS spectrum [M]⁺ =292.1679 (C₁₇ H₂₄ O₄); Calcd. value: 292.1675 [α]_(D) ²⁶ =+2.0° (c=2.0, EtOH)

Referential Example 65 2,2,2-Trichloroethyl 3-(R)-benzyloxycarbonylnonanoate

Following the procedure described in Referential Example 8, but using 3-(R)-benzyloxycarbonylnonanoic acid (2.7 g), prepared in Referential Example 64, and trichloroethanol (1.7 ml), the compound was obtained (3.17 g).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=6.8 Hz), 1.11-1.37 (8H, complex), 1.48-1.78 (2H, complex), 2.60 (1H, dd, J=15.9 and 3.7 Hz), 2.89 (1H, dd, J=15.9 and 9.0 Hz), 2.96 (1H, m), 4.65 (1H, d, J=12.0 Hz), 4.72 (1H, d, J=12.0 Hz), 5.10 (1H, d, J=12.4 Hz), 5.18 (1H, d, J=12.4 Hz), 7.25-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2930 (m), 1758 (s), 1736 (s)

Mass spectrum [M]⁺ =422

High resolution MS spectrum [M]⁺ =422.0826 (C₁₉ H₂₅ O₄ (³⁵ Cl)₃); Calcd. value: 422.0819 [α]_(D) ²⁶ =-0.99° (c=5.9, EtOH)

Referential Example 66 2-(R)-(2,2,2-Trichloroethoxycarbonyl)methyloctanoic acid

Following the procedure described in Referential Example 9, but using 2,2,2-trichloroethyl 3-(R)-benzyloxycarbonylnonanoate (3.12 g), prepared in Referential Example 65, the desired compound (2.39 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.8 Hz), 1.18-1.45 (8H, complex), 1.49-1.81 (2H, complex), 2.61 (1H, dd, J=15.1 and 2.9 Hz), 2.88 (1H, dd, J=15.1 and 9.3 Hz), 2.94 (1H, m), 4.72 (1H, d, J=12.2 Hz), 4.79 (1H, d, J=12.2 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 2930 (m), 1759 (s), 1710 (s)

Mass spectrum [M+H]⁺ =333

High resolution MS spectrum [M+H]⁺ =333.0453 (C₁₂ H₂₀ O₄ (³⁵ Cl₃)₃); Calcd. value: 333.0428 [α]_(D) ²⁶ =-11.17° (c=4.0, EtOH)

Referential Example 67 N¹ -Benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methyloctyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 10, but using an acid chloride of 2-(R)-(2,2,2-trichloroethoxycarbonyl)methyloctanoic acid (464 mg), prepared in Referential Example 66, and (S)-N¹ -benzyloxycarbonylpiperazic acid tert-butyl ester, the desired compound (772 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.84 (3H, t, J=6.6 Hz), 0.90-2.12 (14H, complex), 1.43 (9H, s), 2.60 (1H, dd, J=17.2 and 4.0 Hz), 2.97 (1H, dd, J=17.2 and 9.2 Hz), 3.13 (1H, m), 4.28 (1H, m), 4.61 (1H, d, J=11.9 Hz), 4.78 (1H, d, J=11.9 Hz), 5.13 (1H, d, J=11.9 Hz), 5.21 (1H, d, J=11.9 Hz), 5.27 (1H, dd, J=4.6 and 4.0 Hz), 7.24-7.41 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2931 (m), 1736 (s), 1677 (s)

Mass spectrum [M]⁺ =634

High resolution MS spectrum [M]⁺ =636.1935 (C₂₉ H₄₁ _(O) ₇ N₂ (³⁵ Cl)₂ (³⁷ Cl); Calcd.

value: 636.1950 [α]_(D) ²⁶ =-6.9° (c=2 0, CHCl₃)

Referential Example 68 N¹ -Benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxooctyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 11, but using N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methyloctyl]-(S)-piperazic acid tert-butyl ester (765 mg), prepared in Referential Example 67, the desired compound (495 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.83 (3H, t, J=6.6 Hz), 0.96-2.12 (14H, complex), 1.43 (9H, s), 2.48 (1H, dd, J=17.2 and 3.6 Hz), 2.82 (1H, dd, J=17.2 and 10.6 Hz), 3.08 (1H, m), 3.12 (1H, m), 4.25 (1H, m), 5.13 (1H, d, J=12.2 Hz), 5.20 (1H, d, J=12.2 Hz), 5.27 (1H, dd, J=4.6 and 4.0 Hz), 7.22-7.48 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3186 (w), 2931 (m), 1736 (s), 1678 (s)

Mass spectrum [M+H]⁺ =505

High resolution MS spectrum [M+H]⁺ =505.2913 (C₂₇ H₄₁ O₇ N₂); Calcd. value: 505.2914 [α]_(D) ²⁶ =-23.5° (c=1.0, EtOH)

Referential Example 69 N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 12, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxooctyl]-(S)-piperazic acid tert-butyl ester (485 mg), prepared in Referential Example 68, and O-benzylhydroxylamine, the desired compound (577 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.83 (3H, t, J=6.6 Hz), 0.88-2.08 (15H, complex), 1.42 (9H, s), 2.30 (1H, m), 3.20 (1H, m), 3.43 (1H, m), 4.24 (1H, m), 4.82 (1H, d, J=11.6 Hz), 4.88 (1H, d, J=11.6 Hz), 5.13 (1H, d, J=12.2 Hz), 5.20 (1H, d, J=12.2 Hz), 5.25 (1H, br.t, J=4.0 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 3251 (w), 2931 (m), 1735 (s), 1675 (s)

Mass spectrum [M]⁺ =609

High resolution MS spectrum [M]⁺ =609.3395 (C₃₄ H₄₇ O₇ N₃); Calcd. value: 609.3414 [α]_(D) ²⁶ =-37.8° (c=1.0, CHCl₃)

Referential Example 70 N¹ -Benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid

Following the procedure described in Referential Example 13, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid tert-butyl ester (565 mg), prepared in Referential Example 69, the desired compound (458 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 0.92-2.62 (16H, complex), 2.82-3.29 (2H, complex), 4.10 (1H, m), 4.71-5.40 (5H, complex), 7.00-7.58 (10H, complex), 8.02 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3227 (m), 2930 (s), 1718 (s), 1672 (s), 1608 (s)

Mass spectrum [M-C₆ H₅ ]⁺ =476 [α]_(D) ²⁶ =17.3° (c=1.0, EtOH)

Referential Example 71 4-(S)-Isopropyl-3-(1-oxononyl)-2-oxazolidinone

Following the procedure described in Referential Example 3, but using 4-(S)-isopropyl-2-oxazolidinone (4.99 g) and nonanoyl chloride (7.16 g), the desired compound (10.12 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, d, J=7.3 Hz), 0.88 (3H, t, J=6.8 Hz), 0.92 (3H, d, J=7.3 Hz), 1.18-1.45 (12H, complex), 1.56-1.75 (2H, complex), 2.38 (1H, d,hep, J=3.3 and 7.3 Hz), 2.78-3.07 (2H, complex), 4.20 (1H, dd, J=9.2 and 3.3 Hz), 4.26 (1H, t, J=9.2 Hz), 4.44 (1H, dt, J=7.9 and 3.3 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 2928 (m), 1785 (s), 1703 (s)

Mass spectrum [M]⁺ =269

High resolution MS spectrum [M]⁺ =269.2001 (C₁₅ H₂₇ O₃ N); Calcd. value: 269.1991 [α]_(D) ²⁶ =+67.4° (c=1.0, CHCl₃)

Referential Example 72 4-(S)-Isopropyl-3-[2-(R)-tert-butoxycarbonylmethyl-1-oxononyl]-2-oxazolidinone

Following the procedure described in Referential Example 4, but using 4-(S)-isopropyl-3-(1-oxononyl)-2-oxazolidinone (10.09 g), prepared in Referential Example 71, and tert-butyl bromoacetate (30 ml), the desired compound (12.62 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.9 Hz), 0.91 (3H, d, J=6.6 Hz), 0.93 (3H, d, J=6.6 Hz), 1.17-1.50 (11H, complex), 1.41 (9H, s), 1.60 (1H, m), 2.37 (1H, m), 2.43 (1H, dd, J.=16.5 and 4.0 Hz), 2.74 (1H, dd, J=16.5 and 9.9 Hz), 4.09-4.31 (3H, complex), 4.43 (1H, dt, J=7.9. and 4.0 Hz)

IR absorption spectrum (liquid film, KBr pellet): 2925 (m), 1763 (s), 1731 (s), 1704 (s)

Mass spectrum [M+H]⁺ =384

High resolution MS spectrum [M+H]⁺ =384.2752 (C₂₁ H₃₈ O₅ N); Calcd. value: 384.2750

m.p. 49°-51° (H₂ O--MeOH) [α]_(d) ²⁶ =+48.5° (c=1.0, CHCl₃)

Referential Example 73 tert-Butyl 3-(R)-benzyloxycarbonyldecanoate

Following the procedure described in Referential Example 63, but using 4-(S)-isopropyl-3-[2-(R)-tertbutoxycarbonylmethyl-1-oxononyl)-2-oxazolidinone (12.44 g), prepared in Referential Example 72, the desired compound (10.88 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.14-1.33 (10H, complex), 1.40 (9H, s), 1.49 (1H, m), 1.62 (1H, m), 2.36 (1H, dd, J=16.5 and 6.5 Hz), 2.64 (1H, dd, J=16.5 and 9.2 Hz), 2.84 (1H, m), 5.09 (1H, d, J=12.2 Hz), 5.17 (1H, d, J=12.2 Hz), 7.29-7.40 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2929 (s), 1736 (s)

Mass spectrum [M+H]⁺ =363

High resolution MS spectrum [M+H]⁺ =363.2525 (C₂₂ H₃₅ O₄); Calcd. value: 363.2535

Referential Example 74 3-(R)-Benzyloxycarbonyldecanoic acid

Following the procedure described in Referential Example 7, but using 3-(R)-tert-butyl benzyloxycarbonyldecanoate (10.50 g), prepared in Referential Example 73, the desired compound (8.29 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.10-1.38 (10H, complex), 1.42-1.77 (2H, complex), 2.48 (1H, dd, J=16.2 and 4.6 Hz), 2.79 (1H, dd, J=16.5 and 9.2 Hz), 2.88 (1H, m), 5.12 (1H, d, J=12.5 Hz), 5.17 (1H, d, J=12.5 Hz), 7.25-7.42 (5H, complex) [α]_(D) ²⁶ =+3.0° (c=1.0, EtOH)

Referential Example 75

2,2,2-Trichloroethyl 3-(R)-benzyloxycarbonyldecanoate

Following the procedure described in Referential Example 8, but using 3-(R)-benzyloxycarbonyldecanoic acid (8.26 g), prepared in Referential Example 74, and trichloroethanol (11.5 ml), the desired compound (11.44 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 1.10-1.39 (10H, complex), 1.46-1.82 (2H, complex), 2.60 (1H, dd, J=15.2 and 4.0 Hz), 2.89 (1H, dd, J=15.2 and 9.2 Hz), 2.96 (1H, m), 4.65 (1H, d, J=12.2 Hz), 4.72 (1H, d, J=12.2 Hz), 5.10 (1H, d, J=11.9 Hz), 5.18 (1H, d, J=11.9 Hz), 7.25-7.45 (5H, complex)

Mass spectrum [M]⁺ =436

High resolution MS spectrum [M]⁺ =436.0992 (C₂₀ H₂₇ O₄ (³⁵ Cl)₃); Calcd. value: 436.0974 [α]_(D) ²⁶ =-0.69° (c=4.1, CHCl₃)

Referential Example 76 2-(R)-(2,2,2-Trichloroethoxycarbonyl)methylnonanoic acid

Following the procedure described in Referential Example 9, but using 2,2,2-trichloroethyl 3-(R)-benzyloxycarbonyldecanoate (11.39 g), prepared in Referential Example 65, the desired compound (7.66 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.8 Hz), 1.15-1.45 (10H, complex), 1.50-1.82 (2H, complex), 2.61 (1H, dd, J=15.1 and 3.4 Hz), 2.80-3.01 (2H, complex), 4.72 (1H, d, J=11.7 Hz), 4.79 (1H, d, J=11.7 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 2929 (s), 1759 (s), 1710 (s)

Mass spectrum [M+H]⁺ =347

High resolution MS spectrum [M+H]⁺ =347.0604 (C₁₃ H₂₂ O₄ (³⁵ Cl)₃); Calcd. value: 347.0584 [α]_(D) ²⁶ =+11.8° (c=1.0, EtOH)

Referential Example 77 N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methylnonyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 10, but using an acid chloride of 2-(R)-(2,2,2-trichloroethoxycarbonyl)methylnonanoic acid (376 mg), prepared in Referential Example 76, and (S)-N'-benzyloxycarbonylpiperazic acid tert-butyl ester (349 mg) there was obtained the desired compound (542 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 0.95-1.68 (13H, complex), 1.43 (9H, s), 1.73-2.16 (3H, complex), 2.60 (1H, dd, J=17.2 and 3.6 Hz), 2.94 (1H, dd, J=17.2 and 10.6 Hz), 3.12 (1H, m), 3.43 (1H, m), 4.28 (1H, m), 4.61 (1H, d, J=12.0 Hz), 4.77 (1H, d, J=12.0 Hz), 5.14 (1H, d, J=12.5 Hz), 5.21 (1H, d, J=12.5 Hz), 5.27 (1H, t, J=4.3 Hz), 7.24-7.41 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2930 (s), 1733 (s), 1678 (s)

Mass spectrum [M]⁺ =648

High resolution MS spectrum [M]⁺ =648.2146 (C₃₀ H₄₃ O₇ N₂ (³⁵ Cl)₃); Calcd. value: 648.2136 [α]_(D) ²⁶ =-7.3° (c=1.0, CHCl₃)

Referential Example 78 N¹ -Benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxononyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 11, but using N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methylnonyl]-(S)-piperazic acid tert-butyl ester (541 mg), prepared in Referential Example 77, the desired compound (304 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=7.0 Hz), 0.91-2.11 (16H, complex), 1.43 (9H, s), 2.48 (1H, dd, J=17.2 and 3.3 Hz), 2.81 (1H, dd, J=17.2 and 10.6 Hz), 3.06 (1H, m), 3.32 (1H, br.t, J=11.2 Hz), 4.24 (1H, m), 5.13 (1H, d, J=11.9 Hz), 5.20 (1H, d, J=11.9 Hz), 5.27 (1H, br.t, J=4.0 Hz), 7.23-7.41 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3184 (m), 2929 (s), 1733 (s), 1640 (s)

Mass spectrum [M+H]⁺ =519

High resolution MS spectrum [M+H]⁺ =519.3064 (C₂₈ H₄₃ O₇ N₂) ; Calcd. value: 519.3070

Referential Example 79 N¹ -Benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 12, but using tert-butyl N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxononyl]-(S)-piperazic acid (300 mg), prepared in Referential Example 78, and O-benzylhydroxylamine, the desired compound (347 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, t, J=7.1 Hz), 0.90-2.08 (16H, complex), 1.43 (9H, s), 2.10-2.50 (2H, complex), 3.20 (1H, m), 3.42 (1H, m), 4.25 (1H, m), 4.82 (1H, d, J=11.2 Hz), 4.89 (1H, d, J=11.2 Hz), 5.13 (1H, d, J=12.4 Hz), 5.20 (1H, d, J=12.4 Hz), 5.26 (1H, dd, J=4.4 and 3.4 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 3252 (m), 2929 (s), 1735 (s), 1675 (s)

Mass spectrum [M+H]⁺ =624

High resolution MS spectrum [M+H]⁺ =624.3658 (C₃₅ H₅₀ O₇ N₃); Calcd. value: 624.3648 [α]_(D) ²⁶ =-41.8° (c=1.0, CHCl₃)

Referential Example 80 N¹ -Benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid

Following the procedure described in Referential Example 13, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid tert-butyl ester (339 mg), prepared in Referential Example 79, the desired compound (249 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.89 (3H, t, J=6.7 Hz), 0.92-2.07 (16H, complex), 2.10-2.52 (2H, complex), 2.88-3.22 (2H, complex), 4.10 (1H, m), 4.68-5.33 (5H, complex), 7.00-7.50 (10H, complex), 12.30 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3220 (m), 2928 (s), 1713 (s), 1672 (s), 1601 (s)

Mass spectrum [M-NHOBn]⁺ =445 [α]_(D) ²⁶ =-23.1° (c=1.0, EtOH)

Referential Example 81 4-(S)-Isopropyl-3-(1-oxododecyl)-2-oxazolidinone

Following the procedure described in Referential Example 3, but using 4-(S)-isopropyl-2-oxazolidinone (5.39 g) and dodecanoyl chloride (9.46 g), the desired compound (11.3 g) was obtained as a colorless oil.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, d, J=6.9 Hz), 0.88 (3H, t, J=0.87 ppm and 0.92 ppm, overlapped), 0.92 (3H, d, J=6.9 Hz), 1.17-1.43 (16H, complex), 1.53-1.78 (2H, complex), 2.38 (1H, d,hep, J=3.3 and 6.9 Hz), 2.77-3.07 (2H, complex), 4.20 (1H, dd, J=8.6 and 3.3 Hz), 4.26 (1H, t, J=8.6 Hz), 4.44 (1H, dt, J=8.6 and 3.3 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 2925 (s), 1785 (s), 1704 (s)

Mass Spectrum. [M+H]⁺ =312

High resolution MS spectrum [M+H]⁺ =312.2532 (C₁₈ H₃₄ O₃ N) ; Calcd. value: 312.2539 [α]_(D) ²⁶ =+54.1° (c=1.0, CHCl₃)

Referential Example 82 4-(S)-Isopropyl-3-[2-(R)-tert-butoxycarbonylmethyl-1-oxododecyl]-2-oxazolidinone

Following the procedure described in Referential Example 4, but using 4-(S)-isopropyl-3-(1-oxododecyl)-2-oxazolidone (11.27 g), prepared in Referential Example 81, and tert-butyl bromoacetate (30 ml), the desired compound (14.10 g) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=7.3 Hz), 0.91 (3H, d, J=6.6 Hz), 0.93 (3H, d, J=6.6 Hz), 1.11-1.51 (17H, complex), 1.41 (9H, s), 1.61 (1H, m), 2.37 (1H, m), 2.42 (1H, dd, J=16.6 and 4.4 Hz), 2.74 (1H, dd, J=16.6 and 10.3 Hz), 4.09-4.30 (3H, complex), 4.43 (1H, dt, J=7.8 and 3.7 Hz)

IR absorption spectrum (liquid film kBr pellet) cm⁻¹ : 2922 (s), 1764 (s), 1730 (s), 1700 (s)

Mass spectrum [M+H--tBu]⁺ =369.2505

High resolution MS spectrum [M+H--tBu]⁺ =369.2505 (C₂₀ H₃₅ O₅ N); Calcd. value: 369.2515

m.p. 42°-43° (H₂ O--MeOH) [α]_(D) ²⁶ =+45.0° (c=1.0, CHCl₃)

Referential Example 83 tert-Butyl 3-(R)-benzyloxycarbonyltridecanoate

Following the procedure described in Referential Example 63, but using 4-(S)-isopropyl-3-[2-(R)-tertbutoxycarbonylmethyl-1-oxododecyl)-2-oxazolidinone (13.67 g), prepared in Referential Example 82, the desired compound was obtained (11.92 g).

C₂₅ H₄₀ O₄ (FW=404)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.15-1.71 (18H, complex), 1.40 (9H, s), 2.36 (1H, dd, J=16.2 and 5.3 Hz), 2.64 (1H, dd, J=16.2 and 9.2 Hz), 2.83 (1H, m), 5.09 (1H, d, J=12.5 Hz), 5.17 (1H, d, J=12.5 Hz), 7.27-7.40 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2927 (s), 1734 (s)

Mass spectrum [M+H--tBu]⁺ =348

High resolution MS spectrum [M+H--tBu]⁺ =348.2318 (C₂₁ H₃₂ O₄); Calcd. value: 348.2301 [α]_(D) ²⁶ =+0.76° (c=5.0, CHCl₃)

Referential Example 84 3-(R)-Benzyloxycarbonyltridecanoic acid

Following the procedure described in Referential Example 7, but using tert-butyl 3-(R)-benzyloxycarbonyltridecanoate (11.62 g), prepared in Referential Example 83, the desired compound (9.41 g) was obtained.

C₂₁ H₃₂ O₄ (FW=348)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.12-1.35 (16H, complex), 1.40-1.75 (2H, complex), 2.49 (1H, dd, J=16.1 and 4.4 Hz), 2.79 (1H, dd, J=16.1 and 9.3 Hz), 2.88 (1H, m), 5.12 (1H, d, J=12.7 Hz), 5.17 (1H, d, J=12.7 Hz), 7.25-71.41 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2926 (s), 1737 (s), 1713 (s)

Mass Spectrum [M]⁺ =348

High resolution MS spectrum [M]⁺ =348.2308 (C₂₁ H₃₂ O₄); Calcd. value: 348.2300 [α]_(D) ²⁶ =+2.8° (c=1.0, EtOH)

Referential Example 85 2,2,2-Trichloroethyl 3-(R)-benzyloxycarbonyltridecanoate

Following the procedure described in Referential Example 8, but using 3-(R)-benzyloxycarbonyltridecanoic acid (9.35 g), prepared in Referential Example 84, and trichloroethanol (11.0 ml), the desired compound (10.86 g) was obtained.

C₂₃ H₃₃ O₄ Cl₃ (FW=478, Cl=35)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 1.13-1.39 (16H, complex), 1.48-1.79 (2H, complex), 2.60 (1H, dd, J=15.1 and 3.4 Hz), 2.81-3.02 (2H, complex), 4.65 (1H, d, J=12.0 Hz), 4.72 (1H, d, J=12.0 Hz), 5.10 (1H, d, J=12.4 Hz), 5.18 (1H, d, J=12.4 Hz), 7.28-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2926 (s), 1759 (s), 1737 (s)

Mass spectrum [M]⁺ =478

High resolution MS spectrum [M]⁺ =478.1428 (C₂₃ H₃₃ H₄ (³⁵ Cl)₃); Calcd. value: 478.1445 [α]_(D) ²⁶ =-0.57° (c=3.0, CHCl₃)

Referential Example 86 2-(R)-(2,2,2-Trichloroethoxycarbonyl)methyldodecanoic acid

Following the procedure described in Referential Example 9, but using 2,2,2-trichloroethyl 3-(R)-benzyloxycarbonyltridecanoate (10.78 g), prepared in Referential Example 65, the desired compound (7.01 g) was obtained.

C₁₆ H₂₇ O₄ Cl₃ (FW=388, Cl=35)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.5 Hz), 1.15-1.45 (16H, complex), 1.50-1.82 (2H, complex), 2.61 (1H, dd, J=15.2 and 2.2 Hz), 3.80-3:01 (2H, complex), 4.72 (1H, d, J=11.9 Hz), 4.79 (1H, d, J=11.9 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 2927 (s), 1759 (s), 1710 (s)

Mass spectrum [M+H]⁺ =389

High resolution MS spectrum [M+H]⁺ =389.1036 (C₁₆ H₂₈ O₄ (³⁵ Cl)₃); Calcd. value: 389.1053 [α]_(D) ²⁶ =+11.7° (c=1.0, EtOH)

Referential Example 87 N¹ -Benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methyldodecyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 10, but using an acid chloride of 2-(R)-(2,2,2-trichloroethoxycarbonyl)methyldodecanoic acid (329 mg), prepared in Referential Example 66, and -(S)-N'-benzyloxycarbonylpiperazic acid tert-butyl ester desired compound (492 mg) was obtained.

C₃₃ H₄₉ O₇ N₂ Cl₃ (FW=690, Cl=35)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 0.94-1.39 (18H, complex), 1.43 (9H, s), 1.49-1.71 (2H, complex), 1.78-2.13 (2H, complex), 2.60 (1H, dd, J=17.5 and 3.6 Hz), 2.94 (1H, dd, J=17.5 and 10.9 Hz), 3.14 (1H, m), 3.43 (1H, m), 4.27 (1H, m), 4.61 (1H, d, J=11.9 Hz), 4.77 (1H, d, J=11.9 Hz), 5.14 (1H, d, J=11.9 Hz), 5.21 (1H, d, J=11.9 Hz), 5.27 (1H, t, J=4.0 Hz), 7.23-7.41 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2927 (s), 1740 (s), 1677 (s)

Mass spectrum [M]⁺ =690

High resolution MS spectrum [M]⁺ =690.2598 (C₃₃ H₄₉ O₇ N₂ (³⁵ Cl)₃); Calcd. value: 690.2606 [α]_(D) ²⁶ =-7.0° (c=2.0, CHCl₃)

Referential Example 88 N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxododecyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 11, but using N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methyldodecyl]-(S)-piperazic acid tert-butyl ester (489 mg), prepared in Referential Example 87, there was obtained the desired compound (407 mg).

C₃₁ H₄₈ O₇ N₂ (FW=560)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 0.94-2.11 (22H, complex), 1.43 (9H, s), 2.48 (1H, dd, J=17.5 and 3.6 Hz), 2.82 (1H, dd, J=17.5 and 10.9 Hz), 3.07 (1H, m), 3.31 (1H, br.t, J=10.1 Hz), 4.25 (1H, m), 5.13 (1H, d, J=11.8 Hz), 5.20 (1H, d, J=11.8 Hz), 5.27 (1H, dd, J=5.3 and 3.3 Hz), 7.23-7.40 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3185 (w), 2927 (s), 1.736 (s), 1678 (s)

Mass spectrum [M-tBuO]⁺ =487

High resolution MS spectrum [M-tBuO]⁺ =487.2820 (C₂₇ H₃₉ O₆ N₂); Calcd. value: 487.2808 [α]_(D) ²⁶ =20.5° (c=1.0, EtOH)

Referential Example 89 N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 12, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxododecyl]-(S)-piperazic acid tert-butyl ester(403 mg), prepared in Referential Example 88, and O-benzylhydroxylamine, the desired compound was obtained (463 mg).

C₃₈ H₅₅ O₇ N₃ (FW=665)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t, J=6.6 Hz), 0.94-2.45 (24H, complex), 1.42 (9H, s), 3.21 (1H, m), 3.46 (1H, m), 4.23 (1H, m), 4.82 (1H, d, J=11.2 Hz), 4.87 (1H, d, J=11.2 Hz), 5.13 (1H, d, J=12.5 Hz), 5.20 (1H, d, J=12.5 Hz), 5.24 (1H, m), 7.20-7.49 (10H, complex), 8.41 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3252 (w), 2927 (s), 1733 (s), 1675 (s)

Mass spectrum [M+H]⁺ =666

High resolution MS spectrum [M+H]⁺ =666.4136 (C₃₆ H₅₆ O₇ N₃); Calcd. value: 666.4118 [α]_(D) ²⁶ =-36.2° (c=1.0, CHCl₃)

Referential Example 90 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid

Following the procedure described in Referential Example 13, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid tert-butyl ester (457 mg), prepared in Referential Example 89, the desired compound (370 mg) was obtained.

C₃₄ H₄₇ O₇ N₃ (FW=609)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.88 (3H, t), 0.96-1.38 (16H, complex), 1.38-1.69 (3H, complex), 1.82-2.06 (2H, complex), 2.21-2.59 (2H, complex), 2.90-3.18 (3H, complex), 4.11 (1H, br.d, J=12.5 Hz), 4.85 (1H, br.s), 4.89-5.08 (3H, complex), 5.17 (1H, d, J=11.9 Hz), 7.02-7.51 (11H, complex), 12.32 (1H, s)

IR absorption spectrum (liquid kBr pellet): 3231 (w), 2926 (s), 1710 (s), 1672 (s), 1604 (s) [α]_(D) ²⁶ =-28.1° (c=1.0, EtOH)

Referential Example 91 4-(S)-Isopropyl-3-(1-oxo-4-phenylbutyl)-2-oxazolidinone

Following the procedure described in Referential Example 3, but using 4-(S)-isopropyl-2-oxazolidinone (6.88 g) and 4-phenylbutyryl chloride (6.88 g), the desired compound (10.23 g) was obtained as a colorless oil.

C₁₆ H₂₁ O₃ N (FW=275)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.86 (3H, d, J=6.6 Hz), 0.91 (3H, d, J=6.6 Hz), 1.85-2.21 (2H, complex), 2.36 (1H, d,hep, J=6.6 and 4.0 Hz), 2.69 (2H, t, J=7.9 Hz), 2.91 (1H, dt, J=17.2.and 7.3 Hz), 3.02 (1H, dr, J=17.2 and 7.3 Hz), 4.18 (1H, dd, J=8.6 and 4.0 Hz), 4.24 (1H, t, J=8.6 Hz), 4.41 (1H, dt, J=8.6 and 4.0 Hz), 7.12.-7.36 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2964 (m), 1781 (s), 1701 (s)

Mass spectrum [M]⁺ =275

High resolution MS spectrum [M]⁺ =275.1507 (C₁₆ H₂₁ O₃ N); Calcd. value: 275.1522 [α]_(D) ²⁶ =+60.6° (c=1.0, CHCl₃)

Referential Example 92 4-(S)-Isopropyl-3-[2-(R)-tert-butoxycarbonylmethyl-1-oxo-4-phenylbutyl]-2-oxazolidinone

Following the procedure described in Referential Example 4, but using 4-(S)-isopropyl-3-(1-oxo-4-phenylbutyl)-2-oxazolidinone (8.83 g), prepared in Referential Example 91, and tert-butyl bromoacetate (25.0 ml), there was obtained the desired compound (7.35 g).

C₂₂ H₃₁ NO₅ (FW=389)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.89 (3H, d, J=6.9 Hz), 0.91 (3H, d, J=6.9 Hz), 1.42 (9H, s), 1.78 (1H, m), 2.01 (1H, m), 2.35 (1H, m), 2.47 (1H, dd, J=16.5 and 4.6 Hz), 2.57-2.74 (2H, complex), 2.81 (1H, dd, J=16.5 and 9.9 Hz), 4.08-4.37 (4H, complex), 7.11-7.32 (5H complex)

IR absorption spectrum (liquid kBr pellet): 2978 (w), 1767 (s), 1730 (s), 1691 (s)

Mass spectrum [M]⁺ =389

High resolution MS spectrum [M]⁺ =389.2208 (C₂₂ H₃₁ O₅ N); Calcd. value: 389.2202

m.p. 64°-66° (H₂ O--MeOH) [α]_(D) ²⁶ =+51.6° (c=1.0, CHCl₃)

Referential Example 93 tert-Butyl 3-(R)-benzyloxycarbonyl-5-phenylpentanoate

Following the procedure described in Referential Example 63, using 4-(S)-isopropyl-3-[2-(R)-tert-butoxycarbonylmethyl-1-oxo-4-phenylbutyl)-2-oxazolidinone (5.11 g), prepared in Referential Example 92, the desired compound (4.45 g) was obtained.

C₂₃ H₂₈ O₄ (FW=368)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.40 (9H, s), 1.83 (1H, m), 1.97 (1H, m), 2.41 (1H, dd, J=16.5 and 3.3 Hz), 2.52-2.64 (2H, complex), 2.69 (1H, dd, J=16.5 and 8.6 Hz), 2.89 (1H, m), 5.11 (1H, d, J=12.2 Hz), 5.19 (1H, d, J=12.2 Hz), 7.06-7.44 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2978 (m), 1731 (s), 1604 (w)

Mass spectrum [M]⁺ =368

High resolution MS spectrum [M]⁺ =368.1997 (C₂₃ H₂₈ O₄); Calcd. value: 368.1988 [α]_(D) ²⁶ =+13.0° (c=4.2, CHCl₃)

Referential Example 94 3-(R)-Benzyloxycarbonyl-5-phenylpentanoic acid

Following the procedure described in Referential Example 7, but using tert-butyl 3-(R)-benzyloxycarbonyl-5-phenylpentanoate (4.40 g), prepared in Referential Example 93, the desired compound (3.17 g) was obtained.

C₁₉ H₂₀ O₄ (FW=312)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.77-2.10 (2H, complex), 2.53 (1H, dd, J=15.5 and 3.6 Hz), 2.55-2.70 (2H, complex), 2.83 (1H, dd, J=15.5 and 9.2 Hz), 2.92 (1H, m), 5.12 (1H, d, J=12.5 Hz), 5.18 (1H, d, J=12.5 Hz), 7.06-7.42 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3031 (m), 1736 (s), 1710 (s), 1604 (w)

Mass spectrum [M+H]⁺ =313

High resolution MS spectrum [M+H]⁺ =313.1462 (C₁₉ H₂₁ O₄); Calcd. value: 313.1440 [α]_(D) ²⁶ =+16.0° (c=1.4, EtOH)

Referential Example 95 2,2,2-Trichloroethyl 3-(R)-benzyloxycarbonyl-5-phenylpentanoate

Following the procedure described in Referential Example 8, but using 3-(R)-benzyloxycarbonyl-5-phenylpentanoic acid (3.13 g), prepared in Referential Example 94, and trichloroethanol (4.3 ml), the desired compound (4.07 g) was obtained.

C₂₁ H₂₁ O₄ Cl₃ (FW=442, Cl=35)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.80-2.12 (2H, complex), 2.51-2.83 (3H, complex), 2.88-3.09 (2H, complex), 4.66 (1H, d, J=11.9 Hz), 4.71 (1H, d, J=11.9 Hz), 5.12 (1H, d, J=12.5 Hz), 5.19 (1H, d, J=12.5 Hz), 7.04-7.45 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2958 (w), 2947 (w), 1754 (s), 1733 (s)

Mass spectrum [M]⁺ =442

High resolution MS spectrum [M]⁺ =442.0498 (C₂₁ H₂₁ O₄ (³⁵ Cl)₃); Calcd. value: 442.0505 [α]_(D) ²⁶ =+9.8° (c=3.1, CHCl₃)

Referential Example 96 2-(R)-(2,2,2-Trichloroethoxycarbonyl)methyl-4-phenylbutyric acid

Following the procedure described in Referential Example 9, but using 2,2,2-trichloroethyl 3-(R)-benzyloxycarbonyl-5-phenylpentanoate (4.01 g), prepared in Referential Example 95, the desired compound (2.54 g) was obtained.

C₁₄ H₁₅ O₄ Cl₃ (FW=352, Cl=35)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.92 (1H, m), 2.10 (1H, m), 2.56-2.80 (3H, complex), 2.83-3.08 (2H, complex), 4.72 (1H, d, J=12.2 Hz), 4.78 (1H, d, J=12.2 Hz), 7.11-7.38 (5H, complex)

IR absorption spectrum (liquid film, kBr pellet) cm⁻¹ : 3221 (m), 2951 (m), 1754 (s), 1731 (s), 1694 (s)

Mass spectrum [M]⁺ =352

High resolution MS spectrum [M]⁺ =352.0013 (C₁₄ H₁₅ O₄ (³⁵ Cl)₃); Calcd. value: 352.0035

m.p. 59°-61° C. (H₂ O--MeOH) [α]_(D) ²⁶ =+21.1° (c=17, EtOH)

Referential Example 97 N¹ -Benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methyl-4-phenylbutyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 10, but using an acid chloride of 2-(R)-(2,2,2-trichloroethoxycarbonyl)methyl-4-phenylbutyric acid (353 mg), prepared in Referential Example 96, and (S)-N¹ -benzyloxycarbonylpiperazic acid tert-butyl ester (8 mg), the desired compound (492 mg) was obtained.

C₃₁ H₃₇ O₇ Cl₃ (FW=654)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.42 (9H, s), 1.47-2.19 (6H, complex), 2.31-2.60 (2H, complex), 2.65 (1H, dd, J=17.2 and 4.0 Hz), 3.02 (1H, dd, J=17.2 and 10.6 Hz), 3.22 (1H, m), 3.38 (1H, m), 4.21 (1H, m), 4.63 (1H, d, J=11.9 Hz), 4.78 (1H, d, J=11.9 Hz), 5.16 (2H, s), 5.27 (1H, m), 7.05-7.42 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2955 (m), 1735 (s), 1676 (s)

Mass spectrum [M+H]⁺ =655

High resolution MS spectrum [M+H]⁺ =655.1721 (C₃₁ H₃₈ O₇ N₂ (³⁵ Cl)₃); Calcd. value: 655.1745 [α]_(D) ²⁶ =-16.4° (c=1.0, CHCl₃)

Referential Example 98 N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 11, but using N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methyl-4-phenylbutyl]-(S)-piperazic acid tert-butyl ester (330 mg) prepared in Referential Example 97, there was obtained the desired compound (231 mg).

C₂₉ H₃₆ O₇ N₂ (F=524)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.42 (9H, s), 1.43-2.19 (6H, complex), 2.27-2.73 (2H, complex), 2.52 (1H, dd, J=17.2 and 3.6 Hz), 2.88 (1H, dd, J=17.2 and 10.6 Hz), 3.02-3.47 (2H, complex), 4.20 (1H, m), 5.15 (2H, s), 5.26 (1H, br.dd, J=4.6 and 3.3 Hz), 6.79-7.41 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2977 (m), 1733 (s), 1676 (s)

Mass spectrum [M]⁺ =524

High resolution MS spectrum [M+H]⁺ =525.2593 (C₂₉ H₃₇ O₇ N₂); Calcd. value: 525.2601 [α]_(D) ²⁶ =-29.5° (c=1.0, EtOH)

Referential Example 99 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 12, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-carboxymethyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid tert-butyl ester (228 mg), prepared in Referential Example 98, and O-benzylhydroxylamine, the desired compound (260 mg) was obtained.

C₃₆ H₄₃ O₇ N₃ (FW=629)

NMR spectrum (270 MHz, CDCl₃) δ ppm: 1.30-2.78 (10H, complex), 1.42 (9H, s), 3.14-3.54 (2H, complex), 4.24 (1H, m), 4.83 (1H, d, J=11.7 Hz), 4.88 (1H, d, J=11.7 Hz), 5.15 (2H, s), 5.26 (1H, m), 7.05-7.48 (15H, complex), 8.15 (1H, m)

IR absorption spectrum (liquid film) cm⁻¹ : 3252 (w), 2977 (m), 1732 (s), 1672 (s)

Mass spectrum [M+H]⁺ =630

High resolution MS spectrum [M+H]⁺ =630.3162 (C₃₆ H₄₄ O₇ N₃); Calcd. value: 630.3179

Referential Example 100 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 13, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid tert-butyl ester (260 mg), prepared in Referential Example 99, the desired compound (230 mg) was obtained, containing impurities but which could be used in the following reaction (Example 20) without further purification.

Referential Example 101 N¹ -Benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-4-methyl-1-oxopentyl]-(S)-piperazic acid

Following the procedure described in Referential Example 13, but using N¹ -benzyloxycarbonyl-N² -[2-(R)-benzyloxyaminocarbonyl)methyl-4-methyl-1-oxopentyl]-(S)-piperazic acid tert-butyl ester (401 mg), prepared in Referential Example 46, the desired compound (375 mg) was obtained, having a small amount of impurities, but which was able to be used in the following reactions (Examples 32 and 36) without further purification.

Referential Example 102 2-(S)-(2,2,2-Trichloroethoxycarbonyl)methylheptanoic acid

Following the synthetic procedures (Referential Example 3, 4, 5, 6, 7, 8 and 9) of the corresponding compounds having R-configuration using 4-(R)-isopropyl-2-oxazolidinone and heptanoyl chloride as starting materials, the desired compound was synthesized.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.89 (3H, t, J=6.5 Hz), 1.18-1.47 (6H, complex), 1.47-1.82 (2H, complex), 2.61 (1H, dd, J=15.2 and 2.9 Hz), 2.88 (1H, dd, J=15.2 and 9.3 Hz), 2.94 (1H, m), 4.72 (1H, d, J=12.0 Hz), 4.79 (1H, d, J=12.0 Hz)

IR absorption spectrum (liquid film) cm⁻¹ : 1758 (s), 1709 (s)

High resolution MS spectrum [M+H]⁺ =319.0261 (C₁₁ H₁₈ O₄ Cl₃); Calcd. value: 319.0271 [α]_(D) ²⁶ =-11.2 (c=3.96)

Referential Example 103 N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(S)-(2,2,2-trichloroethoxycarbonyl)methylheptyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 10, but using an acid chloride of 2-(S)-(2,2,2-trichloroethoxycarbonyl)methylheptanoic acid (413 mg), prepared in Referential Example 102 and (S)-N¹ -benzyloxycarbonylpiperazic acid tert-butyl ester (420 mg), the desired compound (675 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.85 (3H, t, J=6.6 Hz), 0.97-2.14 (12H, complex), 1.43 (3H, s), 2.50 (1H, dd, J=17.5 and 4.9 Hz), 2.95 (1H, dd, J=17.5 and 9.9 Hz), 2.97 (1H, m), 3.28 (1H, m), 4.40 (1H, m), 4.58 (1H, d, J=12.5 Hz), 4.85 (1H, d, J=12.5 Hz), 5.10 (1H, d, J=12.5 Hz), 5.21 (1H, d, J=12.5 Hz), 5.29 (1H, br.d, J=4.3 Hz), 7.22-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2931 (m), 1735 (s), 1677 (s)

High resolution MS spectrum [M]⁺ =620.1833 (C₂₈ H₃₉ N₂ O₇ ³⁵ Cl₃); Calcd. value: 620.1823

Referential Example 104 N¹ -benzyloxycarbonyl-N² -[2-(S)-carboxymethyl-1-oxoheptyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 11, but using N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(S)-(2,2,2-trichloroethoxycarbonyl)methylheptyl]-(S)-piperazic acid tert-butyl ester (670 mg), prepared in Referential Example 103, the desired compound (489 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.84 (3H, t, J=6.6 Hz), 0.94-2.17 (12H, complex), 1.42 (9H, s), 2.37 (1H, m), 2.80-3.09 (2H, complex), 3.18 (1H, m), 4.39 (1H, m), 5.00-5.36 (3H, complex), 7.18-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3190 (w), 2932 (s), 1735 (s), 1679 (s)

High resolution MS spectrum [M+H--H₂ O]⁺ =473.2672 (C₂₆ H₃₇ N₂ O₆); Calcd. value: 473.2652

Referential Example 105 N¹ -benzyloxycarbonyl-N² -[2-(S)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 12, but using N¹ -benzyloxycarbonyl-N² -[2-(S)-carboxymethyl-1-oxoheptyl]-(S)-piperazic acid tert-butyl ester(489 mg), prepared in Referential Example 104, and O-benzylhydroxylamine, the desired compound (432 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, t, J=6.6 Hz), 0.93-2.38 (14H, complex), 1.42 (9H, s), 2.81-3.32 (2H, complex), 4.32 (1H, m), 4.75-4.95 (2H, complex), 5.10 (1H, d, J=11.9 Hz), 5.20 (1H, d, J=11.9 Hz), 5.27 (1H, m), 7.22-7.46. (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3252 (m), 2931 (s), 1735 (s), 1675 (s)

High resolution MS spectrum [M+H]⁺ =596.3327 (C₃₃ H₄₆ N₃ O₇); Calcd. value: 596.3335 [α]_(D) ²⁶ =+37.1° (c=1.00, EtOH)

Referential Example 106 N¹ -Benzyloxycarbonyl-N² -[2-(S)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid

Following the procedure described in Referential Example 13, but using N¹ -benzyloxycarbonyl-N² -[2-(S)-(benzyloxycarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid tert-butyl ester (377 mg), prepared in Referential Example 105, there was obtained the desired compound (301 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.83 (3H, t, J=5.8 Hz), 0.97-2.20 (15H, complex), 3.14 (1H, m), 4.24 (1H, m), 4.70-4.95 (2H, br.s), 5.02-5.33 (3H, complex), 7.18-7.48 (10H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 3230 (w), 2940 (m), 1720 (s), 1655 (s) [α]^(D) ²⁶ =+21.4° (c=1.0, EtOH)

Referential Example 107 N¹ -benzyloxycarbonyl-N² -[1-oxo-2-(R)-(2,2,2-trichloroethoxycarbonyl)methyl-4-methylpentyl]-(S)-piperazic acid tert-butyl ester

Following the procedure described in Referential Example 10, but using 2-(R)-(2,2,2-trichloroethoxycarbonyl)methyl-4-methylpentanoic acid (405 mg), prepared in Referential Example 44, and (S)-N¹ -benzyloxycarbonylpiperazic acid tert-butyl ester (421 mg) as starting materials, the desired compound (632 mg) was obtained.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.87 (3H, d, J=6.6 Hz), 0.94 (3H, d, J=6.6 Hz), 1.10-1.71 (4H, complex), 1.43 (9H, s), 1.73-2.15 (3H, complex), 2.62 (1H, dd, J=17.2 and 3.8 Hz), 2.91 (1H, dd, J=17.2 and 10.9 Hz), 3.23 (1H, dd, J=10.5 and 7.7 Hz), 3.47 (1H, dd, J=12.2 and 9.7 Hz), 4.28 (1H, m), 4.61 (1H, d, J=12.5 Hz), 4.77 (1H, d, J=12.5 Hz), 5.14 (1H, d, J=12.5 H), 5.21 (1H, d, J=12.5 Hz), 5.26 (1H, t, J=4.3 Hz), 7.27-7.42 (5H, complex)

IR absorption spectrum (liquid film) cm⁻¹ : 2960 (m), 1735 (s), 1675 (s)

High resolution MS spectrum [M]⁺ =606.1664 (C₂₇ H₃₇ N₂ O₇ ³⁵ Cl₃); Calcd. value: 606.1667 [α]_(D) ²⁶ =-4.3° (c=1.00, CHCl₃)

Referential Example 108 N² -[2-[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl-(S)-piperazic acid (4S,5S)-5-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Referential Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (4S,5S)-5-methyl-3-oxoheptan-4-ylamide (64 mg), prepared in Referential Example 17, were removed by catalytic reduction. The product was purified by preparative reverse phase thin layer chromatography through silica gel (20×20 cm size, 0.25 mm thick), using a 4:6 mixture of water and methanol as a developing solvent and methanol as an eluent, to give the desired compound (35 mg), having a m.p. of 69°-72° C. after recrystallization from a mixture of hexane and ethyl° acetate.

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.85 (3H, t, J=6.7 Hz), 0.87 (3H, t, J=6.1 Hz), 0.92 (3H, d, J=6.7 Hz), 1.00-2.00 (15H, complex), 1.09 (3H, t, J=7.3 Hz), 2.31 (1H, dd, J=12.0 and 4.4 Hz), 2.49 (1H, br.t, J=12.0 Hz), 2.55 (2H, q, J=7.3 Hz), 2.83 (1H, m), 3.01 (1H, br.d, J=12.8 Hz), 3.95 (1H, m), 4.64 (1H, dd, J=8.5 and 4.9 Hz), 4.75 (1H, br.d, J=12.8 Hz), 5.31 (1H, br.s), 7.38 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3303 (m), 1714 (m), 1667 (s), 1626 (s)

High resolution MS spectrum [M+2H--H₂ O]⁺ =424.3072 (C₂₂ H₄₀ N₄ O₄); Calcd. value: 424.3055 [α]_(D) ²⁶ =-30.7° (c=1.01, EtOH)

Referential Example 109 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (4R,5R)-5-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Referential Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid (4S,5S)-5-methyl-3-oxoheptan-4-yalmide (37 mg), prepared in Referential Example 20, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 15:1 mixture of chloroform and methanol as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (19 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.77-0.93 (6H, complex), 0.97 (3H, d, J=6.6 Hz), 1.06 (3H, t, J=7.3 Hz), 1.15-2.63 (17H, complex), 2.52 (2H, q), 2.7014 3.19 (2H, complex), 3.95 (1H, m), 4.47-4.73 (2H, complex), 5.25 (1H, s), 6.81 (1H, d, J=8.6 Hz), 7.70-8.80 (1H, br.s), 9.54 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3274 (m), 2933 (s), 1718 (m), 1665 (s), 1628 (s)

Mass spectrum m/z [M]⁺ =440

High resolution MS spectrum [M-H₃ NO]⁺ =407.2791 (C₂₂ H₃₇ N₃ O₄); Calcd. value: 407.2784 [α]_(D) ²⁶ =+2.7° (c=0.99, EtOH)

Referential Example 110 N² -[2-(R)-(Hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid (4S,5S)-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Referential Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-(benzyloxyaminocarbonyl)methyl-1-oxoheptyl]-(R)-piperazic acid (4R,5R)-5-methyl-3-oxoheptan-4-ylamide (67 mg), prepared in Referential Example 31, were removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 20:1 mixture of chloroform and methanol twice as a developing solvent and a 10:1 mixture of ethyl acetate and methanol as an eluent to, give the desired compound (41 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.80-0.92 (6H, complex), 0.95 (3H, d, J=6.6 Hz), 1.06 (3H, t, J=7.3 Hz), 1.10-2.87 (15H, complex), 2.59 (2H, q, J=7.3 Hz), 3.79 (1H, br.d, J=13.9 Hz), 4.02 (1H, m), 4.35 (1H, br.d, J=12.5 Hz), 4.62 (1H, dd, J=8.2, 5.6 Hz), 5.29 (1H, d, J=3.3 Hz), 7.64 (1H, d, J=17.9 Hz), 9.43 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3270 (s), 2933 (s), 1716 (s), 1650 (s), 1630 (s)

High resolution MS spectrum [M]⁺ =440.3032 (C₂₂ H₄₀ N₄ O₅); Calcd. value: 440.3299 [ ]_(D) ²⁶ =+47.2° (c=1.00, EtOH)

Referential Example 111 [2-[2-(R)-(2-Hydroxyamino-2-oxoethyl)-1-oxoheptyl]-(R)-piperazic acid (4R,5R)-5-methyl-3-oxoheptan-4-ylamide

Following the procedure described in Referential Example 1, the protecting groups of N¹ -benzyloxycarbonyl-N² -[2-(R)-(2-benzyloxyamino-2-oxoethyl)-1-oxoheptyl]-(R)-piperazic acid (4R,5R)-5-methyl-3-oxoheptan-4-ylamide (54 mg), prepared in Referential Example 32, was removed by catalytic reduction. The product was purified by preparative thin layer chromatography through silica gel (20×20 cm size, 0.5 mm thick), using a 25:1 mixture of chloroform and methanol as a developing solvent three times, and a 10:1 mixture of ethyl acetate and methanol as an eluent, to give the desired compound (29 mg).

NMR spectrum (270 MHz, CDCl₃) δ ppm: 0.78-0.99 (9H, complex), 1.03 (3H, t, J=7.3 Hz), 1.11-1.84 (13H, complex), 2.18 (1H, m), 2.30-2.88 (6H, complex), 3.10 (1H, br.d, J=13.9 Hz), 3.82 (1H, d, J=12.5 Hz), 4.19 (1H, m), 4.64 (1H, dd, J=8.6 and 7.9 Hz), 5.23 (1H, d, J=4.0 Hz), 7.18 (1H, br.d, J=8.6 Hz), 7.79 (1H, br.s), 8.84 (1H, br.s)

IR absorption spectrum (liquid film) cm⁻¹ : 3275 (m), 2945 (m), 1715 (m), 1655 (s), 1635 (s)

High resolution MS spectrum [M]⁺ =440.2991 (C₂₂ H₄₀ N₄ O₅); Calcd. value: 440.2998 [α]_(D) ²⁶ =+76.4° (c=0.28, EtOH)

Preparation Example 1

Hard-capsule formulation

100 mg of the powdery compound of Example 3, 150 mg of lactose, 50 mg of cellulose and 6 mg of magnesium stearate were mixed and filled into standard 2-piece hard gelatin capsules. The unit capsules were washed and dried.

Preparation Example 2

Tablet formulation

100 mg of the powdery compound of Example 5, 0.2 mg of colloidal silicon dioxide, 5 mg of magnesium stearate, 275 mg of finely crystalline cellulose, 11 mg of starch and 98.8 mg of lactose were mixed and compressed to appropriate size and weight by conventional means. The tablets, if required, were coated.

Preparation Example 3

Injectable agent formulation

The compound of Example 21 (1.5% by volume) was stirred into propylene glycol (10% by volume) and the mixture was dissolved in water at constant volume for injection and sterilized.

Preparation Example 4

Suspension formulation

100 mg of the powdery compound of Example 34, 100 mg of sodium carboxymethylcellulose, 5 mg of sodium benzoate, 1.0 g of sorbitol (Japanese Pharmacopoeia) and 0.025 ml of vanillin were suspended in 5 ml of a suitable medium. 

We claim:
 1. A compound having the formula: ##STR8## in which: R¹ represents a group of formula:--NR⁴ R⁵ wherein R⁴ and R⁵ are the same or different and each represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms; and R² represents an alkyl group having 6 to 10 carbon atoms.
 2. A compound selected from the following compounds:N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N,N-dimethylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid N,N-dimethylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid N,N-dimethylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N,N-dimethylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid N,N-dimethylamide,and N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid N,N-dimethylamide.
 3. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N-methylamide.
 4. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid N-methylamide.
 5. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid N-methylamide.
 6. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N-methylamide.
 7. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid N-methylamide.
 8. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid N-methylamide.
 9. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N,N-dimethylamide.
 10. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid N,N-dimethylamide.
 11. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid N,N-dimethylamide.
 12. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N,N-dimethylamide.
 13. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid N,N-dimethylamide.
 14. A compound according to claim 2 represented byN² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid N,N-dimethylamide.
 15. An inhibitor of angiogenesis, cancer invasion or cancer metastasis comprising an effective dose of a compound selected from the compounds according to claim 1 in admixture with a pharmaceutically acceptable carrier or vehicle.
 16. An inhibitor of angiogenesis, cancer invasion or cancer metastasis comprising an effective dose of a compound selected from the following compounds in admixture with a pharmaceutically acceptable carrier or vehicle:N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl)-(S)-piperazic acid N-methylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxoheptyl]-(S)-piperazic acid N,N-dimethylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxooctyl]-(S)-piperazic acid N,N-dimethylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxononyl]-(S)-piperazic acid N,N-dimethylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxodecyl]-(S)-piperazic acid N,N-dimethylamide, N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxododecyl]-(S)-piperazic acid N,N-dimethylamide, and N² -[2-(R)-(hydroxyaminocarbonyl)methyl-1-oxo-4-phenylbutyl]-(S)-piperazic acid N,N-dimethylamide. 